LOCUS       sequence-57350-e        3253 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 3253)
  AUTHORS   Jang CW, Magnuson T
  TITLE     A Novel Selection Marker for Efficient DNA Cloning and 
            Recombineering in E. coli.
  JOURNAL   PLoS One. 2013;8(2):e57075. doi: 10.1371/journal.pone.0057075. Epub 
            2013 Feb 20.
  PUBMED    23437314
REFERENCE   2  (bases 1 to 3253)
  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: "PLoS One.";
            date: "2013"; volume: "8(2):e57075. doi"; pages: " 
            10.1371/journal.pone.0057075. Epub 2013 Feb 20"
COMMENT     Sequence Label: pF
FEATURES             Location/Qualifiers
     source          1..3253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      277..732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(364..383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     574..595
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(724..1086)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSKLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWKL"
     primer_bind     858..880
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     872..889
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     873..889
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     896..915
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        896..914
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    923..1030
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     947..963
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     947..963
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(997..1013)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(998..1014)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1043..1061)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1082..1098)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1082..1098)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1095..1117)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1106..1122
                     /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)."
     promoter        complement(join(1130..1136,1137..1154,1155..1160))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1175..1196
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1313..1330)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1484..2072)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1564..1583)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaccctctc cccagagggg cgaggggacc gtattgtgca aatattgtta ccccagcaac
       61 aaacaggctc atacagcccc taaccctttc atggcgatgg ctgggacggt tcagaccttg
      121 ccgaatattc tccagcaccg tctccatgtt ttcaccacaa cagctattcg gctcggtctc
      181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
      241 gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa
      301 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
      361 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
      421 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
      481 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
      541 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
      601 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
      661 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
      721 cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      781 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      841 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata
      901 cgactcacta tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg
      961 atcccccggg ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg
     1021 gcccggtacc cagcttttgt tccctttagt gagggttaat ttcgagcttg gcgtaatcat
     1081 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1141 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1201 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1261 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1321 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1381 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1441 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1501 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1561 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1621 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1681 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1741 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1801 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1861 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1921 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1981 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2041 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2101 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2161 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2221 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggccgcc catctttacc
     2281 aacagaacga ttatttcagt tcgagttcgt tcattgcagc aatgctgaaa ccgccgtcaa
     2341 cgtggaccac ttcaccggag ataccggcag agagatcgga gcacaggaat gccgcagagt
     2401 tacccacatc ttcaatagta acggtacggc gaatcggggt aacggcttcg caatgagcca
     2461 gcattttgcg gaagtctttg ataccggagg ccgccagagt acggatcgga ccagcagaga
     2521 tggcgttaac acgcacacct tccggaccca tcgcgttcgc catatagcgc acgttcgctt
     2581 ccagagacgc ttttgccaga cccataacgt tgtagttcgg gatagcgcgc tcagcgccaa
     2641 ggtaggaaag ggtcagcagg gcagaacccg gattcagcat ggagcggcaa gcttttgcca
     2701 ttgcaacgaa gctgtaggag ctgatgtcgt gggcaatttt gaagccttca cgggtaacgg
     2761 cgttaacata gtcaccatcc agctgatcgc caggtgcaaa accaatagag tgtacgaaac
     2821 cgtcaaattt cggccaaact ttccccagtt cagcgaacat ggtgtcgatg ctggcatctt
     2881 ctgcaacatc gcactgcaga acgatgtcag aacccaattg agcggcaaat tcttctacgc
     2941 ggcctttcag tttgtcgttc tggtaggtga atgccagttc agctccttcg cggtgcatcg
     3001 cctgagcgat accgtaggcg atggatagtt tgctggcaac accggttacc agaatgcgct
     3061 taccggaaag aaaacccata gctttaatcc ttattgttga tgcttgttgt gcctgaaaat
     3121 caggcgattc gtcgttttag taaacagtac gaacagataa acggttatta taatcaacct
     3181 ggctgtgagt agctatagtt gccaggtccg accggagcag gctgcggcag ggggggcgct
     3241 tttcccctca ccc
//
