LOCUS       pET-IDT_N_His_+        5251 bp DNA     circular SYN 11-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5251)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 23, 2024 from SnapGene Viewer 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(92..907)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(92..907)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1029..1617
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1803..1945
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2047..2238)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3047..4129)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4130..4207)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4516..4534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4535..4559
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             4574..4596
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4603..4605
                     /codon_start=1
                     /label=Start Codon
                     /translation="M"
     misc_feature    4614..4659
                     /label=Upstream Flanking Bases
     CDS             4615..4632
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4642..4659
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    4660..4725
                     /label=Downstream Flanking Bases
     terminator      4712..4759
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4796..5224
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
       61 ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa
      121 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
      181 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
      241 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
      301 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
      361 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
      421 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
      481 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
      541 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
      601 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
      661 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
      721 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
      781 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
      841 aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat
      901 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc
      961 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1021 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1081 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1141 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1201 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1261 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1321 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1381 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1441 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1501 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1561 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1621 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1681 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1741 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1801 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt
     1861 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     1921 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     1981 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     2041 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     2101 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     2161 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     2221 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     2281 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg
     2341 gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa
     2401 aatcactcag ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag
     2461 ccagcagcat cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt
     2521 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga
     2581 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc
     2641 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac
     2701 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
     2761 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
     2821 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
     2881 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
     2941 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga
     3001 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt
     3061 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3121 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct
     3181 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc
     3241 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt
     3301 cggtatcgtc gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa
     3361 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga
     3421 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt
     3481 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac
     3541 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca
     3601 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt
     3661 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt
     3721 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt
     3781 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg
     3841 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg
     3901 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg
     3961 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt
     4021 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat
     4081 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc
     4141 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga
     4201 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     4261 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
     4321 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     4381 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     4441 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
     4501 tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
     4561 ctctagaaat aattttgttt aactttaaga aggagatata ccatgggcag cagccatcat
     4621 catcatcatc acagcagcgg cctggtgccg cgcggcagct aacaaagccc gaaaggaagc
     4681 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4741 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     4801 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4861 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4921 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4981 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5041 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5101 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5161 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5221 aattttaaca aaatattaac gcttacaatt t
//