LOCUS       sequence-315036-        2907 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 2907)
  AUTHORS   Hartley RW
  TITLE     Barnase and barstar. Expression of its cloned inhibitor permits 
            expression of a cloned ribonuclease.
  JOURNAL   J Mol Biol 1988 Aug 20;202(4):913-5.
  PUBMED    3050134
REFERENCE   2  (bases 1 to 2907)
  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 Mol 
            Biol"; date: "1988-08-20"; volume: "202(4)"; pages: "913-5"
COMMENT     Sequence Label: pMT416
FEATURES             Location/Qualifiers
     source          1..2907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..17
                     /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     6..28
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     sig_peptide     54..116
                     /codon_start=1
                     /product="signal sequence of E. coli alkaline phosphatase 
                     (Inouye et al., 1982)"
                     /label=phoA signal sequence
                     /translation="MKQSTIALALLPLLFTPVTKA"
     CDS             117..449
                     /codon_start=1
                     /product="ribonuclease from Bacillus amyloliquefaciens"
                     /label=barnase
                     /note="The barnase gene is lethal in standard bacterial 
                     transformation strains."
                     /translation="AQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLADV
                     APGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHYQ
                     TFTKIR"
     CDS             join(1035..1103,1104..1895)
                     /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     complement(1253..1272)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2066..2654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2555..2574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2808..2825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2872..2877,2878..2893,2894..2900)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 ttgtgagcgg ataacaattt cacacaggaa acaggatccg cggagaaaat aaagtgaaac
       61 aaagcactat tgcactggca ctcttaccgt tactgtttac ccctgtgaca aaagccgcac
      121 aggttatcaa cacgtttgac ggggttgcgg attatcttca gacatatcat aagctacctg
      181 ataattacat tacaaaatca gaagcacaag ccctcggctg ggtggcatca aaagggaacc
      241 ttgcagacgt cgctccgggg aaaagcatcg gcggagacat cttctcaaac agggaaggca
      301 aactcccggg caaaagcgga cgaacatggc gtgaagcgga tattaactat acatcaggct
      361 tcagaaattc agaccggatt ctttactcaa gcgactggct gatttacaaa acaacggacc
      421 attatcagac ctttacaaaa atcagataac gaaaaaaacg gcttccctgc gggaggccgt
      481 ttttttcagc tttacataaa gtgtgtaata aatttttctt caaactctga tcggtcaatt
      541 tcactttccg gctctagagc tctagagtcc ggtccaatct gcagccgtcc gagacaggag
      601 gacatcgtcc agctgaaacc ggggcagaat ccggccattt ctgaagagaa aaatggtaaa
      661 ctgatagaat aaaatcataa gaaaggagcc gcacatgaaa aaagcagtca ttaacgggga
      721 acaaatcaga agtatcagcg acctccacca gacattgaaa aaggagcttg cccttccgga
      781 atactacggt gaaaacctgg acgctttatg ggattgtctg accggatggg tggagtaccc
      841 gctcgttttg gaatggaggc agtttgaaca aagcaagcag ctgactgaaa atggcgccga
      901 gagtgtgctt caggttttcc gtgaagcgaa agcggaaggc tgcgacatca ccatcatact
      961 ttcttaatac gatcaatggg agatgaacaa tatggaaaca caaacccgca agcttgcatt
     1021 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1081 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1141 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1201 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1261 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1321 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1381 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1441 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1501 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1561 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1621 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1681 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1741 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1801 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     1861 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     1921 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     1981 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2041 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2101 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2161 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2221 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2281 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2341 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2401 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2461 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2521 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2581 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2641 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2701 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2761 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2821 gcgaggaagc ggaagaggaa ttcgagctcg agcttactcc ccatccccct gttgacaatt
     2881 aatcatcggc tcgtataatg tgtggaa
//
