LOCUS       sequence-254775-        3621 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 3621)
  AUTHORS   Hartley RW, Both V, Hebert EJ, Homerova D, Jucovic M, Nazarov V, 
            Rybajlak I, Sevcik J
  TITLE     Barstar inhibits extracellular ribonucleases of Streptomyces and 
            allows their production from recombinant genes.
  JOURNAL   Protein Peptide Letters 1996 3:225-231
REFERENCE   2  (bases 1 to 3621)
  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: "Protein 
            Peptide Letters 1996 3:225-231"
COMMENT     Sequence Label: pMT1121
FEATURES             Location/Qualifiers
     source          1..3621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..38
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(76..98)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     198..217
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     411..433
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     425..442
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     426..442
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(477..482,483..498,499..505)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    513..529
                     /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)."
     sig_peptide     574..636
                     /codon_start=1
                     /product="signal sequence of E. coli alkaline phosphatase 
                     (Inouye et al., 1982)"
                     /label=phoA signal sequence
                     /translation="MKQSTIALALLPLLFTPVTKA"
     primer_bind     complement(1448..1464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1472..1488
                     /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(1496..1502,1503..1520,1521..1526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1541..1562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1679..1696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1850..2437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1930..1949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2608..3399,3400..3468))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPGGKPSRIIVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3231..3250
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3469..3573)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
       61 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
      121 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
      181 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
      241 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
      301 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      361 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      421 acgttgtaaa acgacggcca gtgaattcga gctcgagctt actccccatc cccctgttga
      481 caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg
      541 aaacaggatc cgcggatccg tggagaaaat aaagtgaaac aaagcactat tgcactggca
      601 ctcttaccgt tactgtttac ccctgtgaca aaagccgacg tctctggtac cgtttgcctg
      661 tctgctctgc cgccggaagc taccgacacc ctgaacctga tcgcttctga cggtccgttc
      721 ccgtactctc aggacggtgt tgttttccag aaccgtgaat ctgttctgcc gacccagtca
      781 tatggttact accacgaata caccgtgatc accccgggtg ctcgtacccg tggtacccgt
      841 cgtatcatca ccggtgaagc tacccaggaa gactactaca ccggtgacca ctacgctacc
      901 ttctctctga tcgatcagac ctgctaacga tttcactttc cggctctaga gctctagagt
      961 ccggtccaat cagcccgcct aatgagcggg cttttttttt gcagccgtcc gagacaggag
     1021 gacatcgtcc agctgaaacc ggggcagaat ccggccattt ctgaagagaa aaatggtaaa
     1081 ctgatagaat aaaatcataa gaaaggagcc gcacatgaaa aaagcagtca ttaacgggga
     1141 acaaatcaga agtatcagcg acctccacca gacattgaaa aaggagcttg cccttccgga
     1201 atactacggt gaaaacctgg acgctttatg ggattgtctg accggatggg tggagtaccc
     1261 gctcgttttg gaatggaggc agtttgaaca aagcaagcag ctgactgaaa atggcgccga
     1321 gagtgtgctt caggttttcc gtgaagcgaa agcggaaggc tgcgacatca ccatcatact
     1381 ttcttaatac gatcaatggg agatgaacaa tatggaaaca cagacccgca agcttggcgt
     1441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaag
     2341 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2401 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2461 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2521 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2581 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2641 agcgatctgt ctatttcgtt catccatagt tgcctgactc cctgtcgtgt agataactat
     2701 gatacgggag ggcttaccac ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2761 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2821 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2881 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2941 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3001 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3061 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3121 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3181 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3241 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3301 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3361 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3421 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3481 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3541 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3601 cgtctaagaa accattatta t
//
