LOCUS       sequence-7053-ex        6239 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 6239)
  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 6239)
  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: pEP PolI
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2818..2835)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2818..2834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2827..2849)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3277..3936)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3851..3870
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3937..4039)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      4384..4606
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4654..5604
                     /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    6130..6151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6166..6171,6172..6189,6190..6196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6204..6220
                     /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     6209..6231
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6228..6239,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6228..6239,1..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 acgctcgcgg 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 ccaaggagtt 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 cagccgacta ctcgcagaat gaactgcgca
     2161 ttatggcgta tctttcgcgt gacaaaggct tgctgaccgc attcgcggaa ggaaaagata
     2221 tccaccgggc aacggccgca gaagtgtttg gtttgccact ggaaaccgtc accagcgagc
     2281 aacgccgtag cgcgaaacgg 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 gaattccact
     2821 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2881 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2941 ttcccaacag ttgcgcagcc tgaatggcga atggcgctaa ccgtttttat caggctctgg
     3001 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     3061 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     3121 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     3181 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
     3241 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
     3301 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac agacggcatg
     3361 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     3421 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     3481 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     3541 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     3601 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     3661 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
     3721 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     3781 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     3841 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     3901 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     3961 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     4021 acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt
     4081 atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta
     4141 ttcggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact
     4201 gaatgtcacg aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc
     4261 agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt
     4321 tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt
     4381 agtgagttat acacagggct gggatctatt ctttttatct ttttttattc tttctttatt
     4441 ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt
     4501 acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata
     4561 cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat
     4621 agtgattaaa atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca
     4681 aatgaactag cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc
     4741 tgtgtggcac tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg
     4801 ttcacttata accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta
     4861 ttagctaaag caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt
     4921 aaaggctttg agattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa
     4981 ttagttttta gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat
     5041 aatctggaac atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta
     5101 ttaaaagaac taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa
     5161 tttaagttca tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg
     5221 gttttgaaac caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat
     5281 aagcgaggcc gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat
     5341 ctcgtaaccg aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc
     5401 attacatcag attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact
     5461 gcaaaaattc agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat
     5521 gatctcaatg gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata
     5581 ctggctaaat acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc
     5641 aagactaaca aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc
     5701 catatgcaca gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt
     5761 tggtgcattc aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt
     5821 aacacctaat agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca
     5881 cctgagacaa cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct
     5941 gcttatcgaa tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa
     6001 atcatggcaa ttctggaaga aatagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6061 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     6121 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     6181 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagct
//
