LOCUS       sequence-7052-ex        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Camps M, Naukkarinen J, Johnson BP, Loeb LA
  TITLE     Targeted gene evolution in Escherichia coli using a highly 
            error-prone DNA polymerase I.
  JOURNAL   Proc Natl Acad Sci U S A. 2003 Aug 19. 100(17):9727-32.
  PUBMED    12909725
REFERENCE   2  (bases 1 to 6238)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2003 Aug 19. 100(17):9727-32."
COMMENT     Sequence Label: pWT PolI
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2817..2834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2817..2833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2826..2848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3276..3935)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3850..3869
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3936..4038)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4383..4605
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4653..5603
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6165..6170,6171..6188,6189..6195)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6203..6219
                     /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)."
     primer_bind     6208..6230
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6227..6238,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6227..6238,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttatggttc agatccccca aaatccactt atccttgtag
       61 atggttcatc ttatctttat cgcgcatatc acgcgtttcc cccgctgact aacagcgcag
      121 gcgagccgac cggtgcgatg tatggtgtcc tcaacatgct gcgcagtctg atcatgcaat
      181 ataaaccgac gcatgcagcg gtggtctttg acgccaaggg aaaaaccttt cgtgatgaac
      241 tgtttgaaca ttacaaatca catcgcccgc caatgccgga cgatctgcgt gcacaaatcg
      301 aacccttgca cgcgatggtt aaagcgatgg gactgccgct gctggcggtt tctggcgtag
      361 aagcggacga cgttatcggt actctggcgc gcgaagccga aaaagccggg cgtccggtgc
      421 tgatcagcac tggcgataaa gatatggcgc agctggtgac gccaaatatt acgcttatca
      481 ataccatgac gaataccatc ctcggaccgg aagaggtggt gaataagtac ggcgtgccgc
      541 cagaactgat catcgatttc ctggcgctga tgggtgactc ctctgataac attcctggcg
      601 taccgggcgt cggtgaaaaa accgcgcagg cattgctgca aggtcttggc ggactggata
      661 cgctgtatgc cgagccagaa aaaattgctg ggttgagctt ccgtggcgcg aaaacaatgg
      721 cagcgaagct cgagcaaaac aaagaagttg cttatctctc ataccagctg gcgacgatta
      781 aaaccgacgt tgaactggag ctgacctgtg aacaactgga agtgcagcaa ccggcagcgg
      841 aagagttgtt ggggctgttc aaaaagtatg agttcaaacg ctggactgct gatgtcgaag
      901 cgggcaaatg gttacaggcc aaaggggcaa aaccagccgc gaagccacag gaaaccagtg
      961 ttgcagacga agcaccagaa gtgacggcaa cggtgatttc ttatgacaac tacgtcacca
     1021 tccttgatga agaaacactg aaagcgtgga ttgcgaagct ggaaaaagcg ccggtatttg
     1081 catttgatac cgaaaccgac agccttgata acatctctgc taacctggtc gggctttctt
     1141 ttgctatcga gccaggcgta gcggcatata ttccggttgc tcatgattat cttgatgcgc
     1201 ccgatcaaat ctctcgcgag cgtgcactcg agttgctaaa accgctgctg gaagatgaaa
     1261 aggcgctgaa ggtcgggcaa aacctgaaat acgatcgcgg tattctggcg aactacggca
     1321 ttgaactgcg tgggattgcg tttgatacca tgctggagtc ctacattctc aatagcgttg
     1381 ccgggcgtca cgatatggac agcctcgcgg aacgttggtt gaagcacaaa accatcactt
     1441 ttgaagagat tgctggtaaa ggcaaaaatc aactgacctt taaccagatt gccctcgaag
     1501 aagccggacg ttacgccgcc gaagatgcag atgtcacctt gcagttgcat ctgaaaatgt
     1561 ggccggatct gcaaaaacac aaagggccgt tgaacgtctt cgagaatatc gaaatgccgc
     1621 tggtgccggt gctttcacgc attgaacgta acggtgtgaa gatcgatccg aaagtgctgc
     1681 acaatcattc tgaagagctc acccttcgtc tggctgagct ggaaaagaaa gcgcatgaaa
     1741 ttgcaggtga ggaatttaac ctttcttcca ccaagcagtt acaaaccatt ctctttgaaa
     1801 aacagggcat taaaccgctg aagaaaacgc cgggtggcgc gccgtcaacg tcggaagagg
     1861 tactggaaga actggcgctg gactatccgt tgccaaaagt gattctggag tatcgtggtc
     1921 tggcgaagct gaaatcgacc tacaccgaca agctgccgct gatgatcaac ccgaaaaccg
     1981 ggcgtgtgca tacctcttat caccaggcag taactgcaac gggacgttta tcgtcaaccg
     2041 atcctaacct gcaaaacatt ccggtgcgta acgaagaagg tcgtcgtata cgccaggcgt
     2101 ttattgcgcc agaggattat gtgattgtct cagcggacta ctcgcagatt gaactgcgca
     2161 ttatggcgca tctttcgcgt gacaaaggct tgctgaccgc attcgcggaa ggaaaagata
     2221 tccaccgggc aacggcggca gaagtgtttg gtttgccact ggaaaccgtc accagcgagc
     2281 aacgccgtag cgcgaaagcg atcaactttg gtctgattta tggcatgagt gctttcggtc
     2341 tggcgcggca attgaacatt ccacgtaaag aagcgcagaa gtacatggac ctttacttcg
     2401 aacgctaccc tggcgtgctg gagtatatgg aacgcacccg tgctcaggcg aaagagcagg
     2461 gctacgttga aacgctggac ggacgccgtc tgtatctgcc ggatatcaaa tccagcaatg
     2521 gtgctcgtcg tgcagcggct gaacgtgcag ccattaacgc gccaatgcag ggaaccgccg
     2581 ccgacattat caaacgggcg atgattgccg ttgatgcgtg gttacaggct gagcaaccgc
     2641 gtgtacgtat gatcatgcag gtacacgatg aactggtatt tgaagttcat aaagatgatg
     2701 ttgatgccgt cgcgaagcag attcatcaac tgatggaaaa ctgtacccgt ctggatgtgc
     2761 cgttgctggt ggaagtgggg agtggcgaaa actgggatca ggcgcactaa gaattcactg
     2821 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2881 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2941 tcccaacagt tgcgcagcct gaatggcgaa tggcgctaac cgtttttatc aggctctggg
     3001 aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact
     3061 ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     3121 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga
     3181 atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg cgtagcacca
     3241 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     3301 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca gacggcatga
     3361 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     3421 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     3481 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     3541 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     3601 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     3661 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
     3721 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     3781 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     3841 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     3901 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     3961 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     4021 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     4081 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     4141 tcggcgcctg tagtgccatt tacccccatt cactgccaga gccgtgagcg cagcgaactg
     4201 aatgtcacga aaaagacagc gactcaggtg cctgatggtc ggagacaaaa ggaatattca
     4261 gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt
     4321 gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta
     4381 gtgagttata cacagggctg ggatctattc tttttatctt tttttattct ttctttattc
     4441 tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta
     4501 cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac
     4561 ccacaactca aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata
     4621 gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa
     4681 atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct
     4741 gtgtggcact actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt
     4801 tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct tatggtgtat
     4861 tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta
     4921 aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat
     4981 tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata
     5041 atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat
     5101 taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat
     5161 ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg
     5221 ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata
     5281 agcgaggccg cccgactgat acgttgattt tccaagttga actagataga caaatggatc
     5341 tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca
     5401 ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg
     5461 caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg
     5521 atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac
     5581 tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca
     5641 agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc
     5701 atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt
     5761 ggtgcattca aagctgttca ccatgaacag atcgacaatg taacagatga acagcatgta
     5821 acacctaata gaacaggtga aaccagtaaa acaaagcaac tagaacatga aattgaacac
     5881 ctgagacaac ttgttacagc tcaacagtca cacatagaca gcctgaaaca ggcgatgctg
     5941 cttatcgaat caaagctgcc gacaacacgg gagccagtga cgcctcccgt ggggaaaaaa
     6001 tcatggcaat tctggaagaa atagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6061 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6121 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     6181 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagct
//
