LOCUS       sequence-7054-ex        3683 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 3683)
  AUTHORS   Shinkai A, Loeb LA
  TITLE     In vivo mutagenesis by Escherichia coli DNA polymerase I. Ile(709) 
            in motif A functions in base selection.
  JOURNAL   J Biol Chem. 2001 Dec 14. 276(50):46759-64.
  PUBMED    11602576
REFERENCE   2  (bases 1 to 3683)
  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: "J Biol 
            Chem. 2001 Dec 14. 276(50):46759-64."
COMMENT     Sequence Label: pLA230
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(19..810,811..879))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHP*TLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     642..661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(880..971)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1037..1625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1117..1136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2514..3329)
                     /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(2514..3329)
                     /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"
     primer_bind     3237..3256
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3321..3340
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     mobile_element  3495..3660
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ggtaccgagc tcggatcctt accaatgctt aatcagtgag gcacctatct cagcgatctg
       61 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
      121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
      181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
      241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
      301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
      361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
      421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
      481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
      541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
      601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
      661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
      721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
      781 atcttttact ttcaccagcg tttatgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
      841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
      901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      961 aaataaacaa ataggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1021 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1081 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1141 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1201 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1261 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1321 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1381 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1441 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     1501 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1561 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1621 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1681 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1741 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1801 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1861 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1921 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1981 cagccggaag ggcgatcaga tctaaactat gacaataaag tcttaaacta gacagaatag
     2041 ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc
     2101 taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg
     2161 ggtcgacgcg gccgctaact ataacggtcc taaggtagcg agtttaaacg atatcggatc
     2221 cggccgccgc tgaggtctgc ctcgtgaaga aggtgttgct gactcatacc aggcctgaat
     2281 cgccccatca tccagccaga aagtgaggga gccacggttg atgagagctt tgttgtaggt
     2341 ggaccagttg gtgattttga acttttgctt tgccacggaa cggtctgcgt tgtcgggaag
     2401 atgcgtgatc tgatccttca actcagcaaa agttcgattt attcaacaaa gccgccgtcc
     2461 cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat taaccaattc tgattagaaa
     2521 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     2581 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     2641 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     2701 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     2761 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     2821 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     2881 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     2941 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     3001 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     3061 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     3121 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     3181 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     3241 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
     3301 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3361 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     3421 tgagacacaa cgtggcttac taggatccga tatcatttaa atctagggat aacagggtaa
     3481 tactagtgtc gaccaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     3541 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     3601 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     3661 ccgtaagatg cttttctgtg act
//
