LOCUS       sequence-169493-        3189 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 3189)
  AUTHORS   McKown RL, Orle KA, Chen T, Craig NL
  TITLE     Sequence requirements of Escherichia coli attTn7, a specific site of
            transposon Tn7 insertion.
  JOURNAL   J Bacteriol 1988 Jan;170(1):352-8.
  PUBMED    2826397
REFERENCE   2  (bases 1 to 3189)
  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 
            Bacteriol"; date: "1988-01"; volume: "170(1)"; pages: "352-8"
COMMENT     Sequence Label: pRM2
FEATURES             Location/Qualifiers
     source          1..3189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(145..249)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     317..335
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(373..395)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     495..514
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     708..730
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     722..739
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     723..739
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1312..1328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1312..1328)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1325..1347)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1336..1352
                     /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(1360..1366,1367..1384,1385..1390))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1405..1426
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1543..1560)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1714..2302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1794..1813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2473..3189,1..75,76..144))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3096..3115
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
       61 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      121 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      181 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      241 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      301 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      361 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      421 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      481 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
      541 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      601 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      661 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      721 ttgtaaaacg acggccagtg ccaagcttgc atgcctgcag gtcgataagg tcgcggcgac
      781 aacagttgcg acggtggtac gcataacttt cataatgtct cctgggagga ttcataaagc
      841 attgtttgtt ggctacgaga agcaaaatag gacaaacagg tgacagttat atgtaaggaa
      901 tatgacagtt ttatgacaga gagataaagt cttcagtctg atttaaataa gcgttgatat
      961 tcagtcaatt acaaacatta ataacgaaga gatgacagaa aaattttcat tctgtgacag
     1021 agaaaaagta gccgaagatg acggtttgtc acatggagtt ggcaggatgt ttgattaaaa
     1081 acataacagg aagaaaaatg ccccgcttac gcagggcatc catttattac tcaaccgtaa
     1141 ccgattttgc caggttacgc ggctggtcaa cgtcggtgcc tttgatcagc gcgacatggt
     1201 aagccagcag ctgcagcgga acggtgtaga agatcggtgc aatcacctct tccacatgcg
     1261 gcatctcgac tctagaggat ccccgggtac cgagctcgaa ttcgtaatca tgtcatagct
     1321 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     1381 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     1441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     1501 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1561 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1621 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     2221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     2341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     2401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2461 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2521 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2581 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2641 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2701 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2761 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2821 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2881 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2941 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3001 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3061 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3121 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3181 gctgttgag
//
