LOCUS       sequence-57351-e        3253 bp    DNA     circular SYN 15-APR-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 Apr 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: pF2
FEATURES             Location/Qualifiers
     source          1..3253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      263..718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(350..369)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     560..581
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(710..1072)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSSKLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWKL"
     primer_bind     844..866
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     858..875
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     859..875
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     882..901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        882..900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    909..1016
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     933..949
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     933..949
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(983..999)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(984..1000)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1029..1047)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1068..1084)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1068..1084)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1081..1103)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1092..1108
                     /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(1116..1122,1123..1140,1141..1146))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1161..1182
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1299..1316)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1470..2058)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1550..1569)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaggggcgag gggaccgtat tgtgcaaata ttgttacccc agcaacaaac aggctcatac
       61 agcccctaac cctttcatgg cgatggctgg gacggttcag accttgccga atattctcca
      121 gcaccgtctc catgttttca ccacaacagc tattcggctc ggtctcaggg ttattgtctc
      181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      241 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      301 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      361 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      421 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      481 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      541 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      601 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      661 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
      721 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      781 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      841 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg
      901 gcgaattgga gctccaccgc ggtggcggcc gctctagaac tagtggatcc cccgggctgc
      961 aggaattcga tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccagc
     1021 ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc atagctgttt
     1081 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     1141 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     1201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1261 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1321 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1381 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1441 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1501 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1561 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1621 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1681 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1741 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1801 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1861 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     1921 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1981 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2041 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2101 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2161 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2221 tctgacagtt accaatgctt aatcagtgag gccgcccatc tttaccaaca gaacgattat
     2281 ttcagttcga gttcgttcat tgcagcaatg ctgaaaccgc cgtcaacgtg gaccacttca
     2341 ccggagatac cggcagagag atcggagcac aggaatgccg cagagttacc cacatcttca
     2401 atagtaacgg tacggcgaat cggggtaacg gcttcgcaat gagccagcat tttgcggaag
     2461 tctttgatac cggaggccgc cagagtacgg atcggaccag cagagatggc gttaacacgc
     2521 acaccttccg gacccatcgc gttcgccata tagcgcacgt tcgcttccag agacgctttt
     2581 gccagaccca taacgttgta gttcgggata gcgcgctcag cgccaaggta ggaaagggtc
     2641 agcagggcag aacccggatt cagcatggag cggcaagctt ttgccattgc aacgaagctg
     2701 taggagctga tgtcgtgggc aattttgaag ccttcacggg taacggcgtt aacatagtca
     2761 ccatccagct gatcgccagg tgcaaaaaca atagagtgta cgaaaccgtc aaatttcggc
     2821 caaactttcc ccagttcagc gaacatggtg tcgatgctgg catcttctgc aacatcgcac
     2881 tgcagaacga tgtcagaacc caattgagcg gcaaattctt ctacgcggcc tttcagtttg
     2941 tcgttctggt aggtgaatgc cagttcagct ccttcgcggt gcatcgcctg agcgataccg
     3001 taggcgatgg atagtttgct ggcaacaccg gttaccagaa tgcgcttacc ggaaagaaaa
     3061 cccatagctt taatccttat tgttgatgct tgttgtgcct gaaaatcagg cgattcgtcg
     3121 ttttagtaaa cagtacgaac agataaacgg ttattataat caacctggct gtgagtagct
     3181 atagttgcca ggtccgaccg gagcaggctg cggcaggggg ggcgcttttc ccctcaccct
     3241 aaccctctcc cca
//
