LOCUS       sequence-254776-        3633 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 3633)
  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 3633)
  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: "Protein 
            Peptide Letters 1996 3:225-231"
COMMENT     Sequence Label: pMT1101
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..68
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(106..128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     228..247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     441..463
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     455..472
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     456..472
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(507..512,513..528,529..535)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    543..559
                     /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     612..674
                     /codon_start=1
                     /product="signal sequence of E. coli alkaline phosphatase 
                     (Inouye et al., 1982)"
                     /label=phoA signal sequence
                     /translation="MKQSTIALALLPLLFTPVTKA"
     primer_bind     complement(1489..1505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1489..1505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    1513..1529
                     /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(1537..1543,1544..1561,1562..1567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1582..1603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1720..1737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1891..2479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1971..1990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2650..3441,3442..3510))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIIVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3273..3292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3511..3615)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
       61 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
      121 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
      181 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      241 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
      301 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      361 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
      421 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga
      481 gctcgagctt actccccatc cccctgttga caattaatca tcggctcgta taatgtgtgg
      541 aattgtgagc ggataacaat ttcacacagg aaacaggatc cgcggatccg cggatccgtg
      601 gagaaaataa agtgaaacaa agcactattg cactggcact cttaccgtta ctgtttaccc
      661 ctgtgacaaa agccgcagac ccggcgctgg ccgacgtctg ccgcaccaag ctgcccagcc
      721 aggcccagga caccctcgcc ctgatcgcca agaacggccc ctacccgtac aaccgggacg
      781 gcgtcgtctt cgagaaccgc gagagccgcc tgccgaagaa gggcaacggc tactaccacg
      841 agttcaccgt ggtcaccccc ggctccaacg accgaggcac ccgacgggtc gtcaccggcg
      901 gctacggcga gcagtactgg tccccggacc actacgcgac cttccaggag atcgacccgc
      961 gctgctaacg atttcacttt ccggctctag agctctagag tccggtccaa tcagcccgcc
     1021 taatgagcgg gctttttttt tgcagccgtc cgagacagga ggacatcgtc cagctgaaac
     1081 cggggcagaa tccggccatt tctgaagaga aaaatggtaa actgatagaa taaaatcata
     1141 agaaaggagc cgcacatgaa aaaagcagtc attaacgggg aacaaatcag aagtatcagc
     1201 gacctccacc agacattgaa aaaggagctt gcccttccgg aatactacgg tgaaaacctg
     1261 gacgctttat gggattgtct gaccggatgg gtggagtacc cgctcgtttt ggaatggagg
     1321 cagtttgaac aaagcaagca gctgactgaa aatggcgccg agagtgtgct tcaggttttc
     1381 cgtgaagcga aagcggaagg ctgcgacatc accatcatac tttcttaata cgatcaatgg
     1441 gagatgaaca atatggaaac acagacccgc aagcttggcg taatcatggt catagctgtt
     1501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1621 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1801 cacagaatca ggggacaacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1981 ggcgtttccc cccggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2701 ttcatccata gttgcctgac tccccgtcgt gtagataact atgatacggg agggcttacc
     2761 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3601 aataggggtt ccgcgcacat ttccccgaaa agt
//
