LOCUS       Exported                8315 bp DNA     circular SYN 23-APR-2025
DEFINITION  Exported.
ACCESSION   V012891
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8315)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8315)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8315)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          5550..5553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             831..2321
                     /codon_start=1
                     /label=AMP-binding protein
                     /translation="MNIPFVVETVLHDGLLKYKFKNSKIRSITTKPGKSKGAIFAYRSK
                     SSMIGGRGVVLTSEEAIQENQDTFTHWTPNVYRYGTYADENRSYTKGHSENNLRQINTF
                     FIDFDIHTAKETISASDILTTAIDLGFMPTMIIKSDKGYQAYFVLETPVYVTSKSEFKS
                     VKAAKIISQNIREYFGKSLPVDLTCNHFGIARIPRTDNVEFFDPNYRYSFKEWQDWSFK
                     QTDNKGFTRSSLTVLSGTEGKKQVDEPWFNLLLHETKFSGEKGLIGRNNVMFTLSLAYF
                     SSGYSIETCEYNMFEFNNRLDQPLEEKEVIKIVRSAYSENYQGANREYITILCKAWVSS
                     DLTSKDLFVRQGWFKFKKKRSERQRVHLSEWKEDLMAYISEKSDVYKPYLVTTKKEIRE
                     VLGIPERTLDKLLKVLKANQEIFFKIKPGRNGGIQLASVKSLLLSIIKVKKEEKESYIK
                     ALTNSFDLEHTFIQETLNKLAERPKTDTQLDLFSYDTG"
     primer_bind     complement(2818..2834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3308..3412
                     /label=AmpR promoter
     CDS             3413..4270
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4444..5032
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5320..5341
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5356..5386
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5394..5410
                     /label=lac operator
                     /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     5418..5434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5455..5473
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5549..5659
                     /label=P59
                     /note="Streptococcus cremoris promoter 59"
     promoter        5550..5553
                     /label=P59
                     /note="Streptococcus cremoris promoter 59"
     misc_feature    5624..5629
                     /label=minus_35_signal
     misc_feature    5647..5652
                     /label=minus_10_signal
     CDS             5737..5817
                     /codon_start=1
                     /label=signal peptide of Usp45
                     /translation="MKKKIISAILMSTVILSAAAPLSGVYA"
     CDS             5863..6303
                     /label=MNase
                     /note="micrococcal nuclease from Staphylococcus aureus"
     terminator      6342..6428
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6447..6474
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             7091..7825
                     /gene="ermBP"
                     /label=rRNA adenine N-6-methyltransferase
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
ORIGIN
        1 aactcaacgc tagtagtgga tttaatccca aatgagccaa cagaaccaga accagaaaca
       61 gaatcagaac aagtaacatt ggatttagaa atggaagaag aaaaaagcaa tgacttcgtg
      121 tgaataatgc acgaaatcgt tgcttatttt ttttaaaagc ggtatactag atataacgaa
      181 acaacgaact gaatagaaac gaaaaaagag ccatgacaca tttataaaat gtttgacgac
      241 attttataaa tgcatagccc gataagattg ccaaaccaac gcttatcagt tagtcagatg
      301 aactcttccc tcgtaagaag ttatttaatt aactttgttt gaagacggta tataaccgta
      361 ctatcattat atagggaaat cagagagttt tcaagtatct aagctactga atttaagaat
      421 tgttaagcaa tcaatcggaa atcgtttgat tgcttttttt gtattcattt atagaaggtg
      481 gagtttgtat gaatcatgat gaatgtaaaa cttatataaa aaatagttta ttggagataa
      541 gaaaattagc aaatatctat acactagaaa cgtttaagaa agagttagaa aagagaaata
      601 tctacttaga aacaaaatca gataagtatt tttcttcgga gggggaagat tatatatata
      661 agttaataga aaataacaaa ataatttatt cgattagtgg aaaaaaattg acttataaag
      721 gaaaaaaatc tttttcaaaa catgcaatat tgaaacagtt gaatgaaaaa gcaaaccaag
      781 ttaattaaac aacctatttt ataggattta taggaaagga gaacagctga atgaatatcc
      841 cttttgttgt agaaactgtg cttcatgacg gcttgttaaa gtacaaattt aaaaatagta
      901 aaattcgctc aatcactacc aagccaggta aaagcaaagg ggctattttt gcgtatcgct
      961 caaaatcaag catgattggc ggtcgtggtg ttgttctgac ttccgaggaa gcgattcaag
     1021 aaaatcaaga tacatttaca cattggacac ccaacgttta tcgttatgga acgtatgcag
     1081 acgaaaaccg ttcatacacg aaaggacatt ctgaaaacaa tttaagacaa atcaatacct
     1141 tctttattga ttttgatatt cacacggcaa aagaaactat ttcagcaagc gatattttaa
     1201 caaccgctat tgatttaggt tttatgccta ctatgattat caaatctgat aaaggttatc
     1261 aagcatattt tgttttagaa acgccagtct atgtgacttc aaaatcagaa tttaaatctg
     1321 tcaaagcagc caaaataatt tcgcaaaata tccgagaata ttttggaaag tctttgccag
     1381 ttgatctaac gtgtaatcat tttggtattg ctcgcatacc aagaacggac aatgtagaat
     1441 tttttgatcc taattaccgt tattctttca aagaatggca agattggtct ttcaaacaaa
     1501 cagataataa gggctttact cgttcaagtc taacggtttt aagcggtaca gaaggcaaaa
     1561 aacaagtaga tgaaccctgg tttaatctct tattgcacga aacgaaattt tcaggagaaa
     1621 agggtttaat agggcgtaat aacgtcatgt ttaccctctc tttagcctac tttagttcag
     1681 gctattcaat cgaaacgtgc gaatataata tgtttgagtt taataatcga ttagatcaac
     1741 ccttagaaga aaaagaagta atcaaaattg ttagaagtgc ctattcagaa aactatcaag
     1801 gggctaatag ggaatacatt accattcttt gcaaagcttg ggtatcaagt gatttaacca
     1861 gtaaagattt atttgtccgt caagggtggt ttaaattcaa gaaaaaaaga agcgaacgtc
     1921 aacgtgttca tttgtcagaa tggaaagaag atttaatggc ttatattagc gaaaaaagcg
     1981 atgtatacaa gccttattta gtgacgacca aaaaagagat tagagaagtg ctaggcattc
     2041 ctgaacggac attagataaa ttgctgaagg tactgaaggc gaatcaggaa attttcttta
     2101 agattaaacc aggaagaaat ggtggcattc aacttgctag tgttaaatca ttgttgctat
     2161 cgatcattaa agtaaaaaaa gaagaaaaag aaagctatat aaaggcgctg acaaattctt
     2221 ttgacttaga gcatacattc attcaagaga ctttaaacaa gctagcagaa cgccctaaaa
     2281 cggacacaca actcgatttg tttagctatg atacaggctg aaaataaaac ccgcactatg
     2341 ccattacatt tatatctatg atacgtgttt gttttttctt tgctgtttag cgaatgatta
     2401 gcagaaatat acagagtaag attttaatta attattaggg ggagaaggag agagtagccc
     2461 gaaaactttt agttggcttg gactgaacga agtgagggaa aggctactaa aacgtcgagg
     2521 ggcagtgaga gcgaagcgaa cacttgattt tttaattttc tatcttttat aggtcattag
     2581 agtatactta tttgtcctat aaactattta gcagcataat agatttattg aataggtcat
     2641 ttaagttgag catattagag gaggaaaatc ttggagaaat atttgaagaa cccgattaca
     2701 tggattggat tagttcttgt ggttacgtgg tttttaacta aaagtagtga atttttgatt
     2761 tttggtgtgt gtgtcttgtt gttagtattt gctagtcaaa gtgattaaat agaattcact
     2821 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2881 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2941 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     3001 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     3061 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3121 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3181 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     3241 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     3301 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3361 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3421 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3481 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3541 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3601 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3661 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3721 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3781 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3841 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3901 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3961 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4021 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4081 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4141 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4201 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4261 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     4321 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4381 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4441 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4501 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4561 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4621 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4681 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4741 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4801 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4861 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4921 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4981 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5041 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5101 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5161 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     5221 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     5281 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     5341 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     5401 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tcgaaattaa
     5461 ccctcactaa agggaacaaa agctggtacc gggccccccc tcgacggtat cgatagcccg
     5521 cctaatgagc gggctttttt ttgatcccct aaccaaagaa gcgcgtaata tcagggttta
     5581 agagacaata agaaaaaaag attgaaaaag tgacattaaa ttcttgacag ggagagatag
     5641 gtttgataga atataatagg ggtaccgagc tcgaattcct gcagcccaaa tcgcgaaacc
     5701 gaacttaatg ggaggaaaaa ttaaaaaaga acagttatga aaaaaaagat tatctcagct
     5761 attttaatgt ctacagtgat actttctgct gcagccccgt tgtcaggtgt ttacgctgac
     5821 acaaactcag atattgctaa acaagatgcg gtcgacccat tgtcaactaa aaaattacat
     5881 aaagaacctg cgactttaat taaagcgatt gatggtgata cggttaaatt aatgtacaaa
     5941 ggtcaaccaa tgacattcag actattattg gttgatacac ctgaaacaaa gcatcctaaa
     6001 aaaggtgtag agaaatatgg tcctgaagca agtgcattta cgaaaaaaat ggtagaaaat
     6061 gcaaagaaaa ttgaagtcga gtttgacaaa ggtcaaagaa ctgataaata tggacgtggc
     6121 ttagcgtata tttatgctga tggaaaaatg gtaaacgaag ctttagttcg tcaaggcttg
     6181 gctaaagttg cttatgttta caaacctaac aatacacatg aacaacattt aagaaaaagt
     6241 gaagcacaag cgaaaaaaga gaaattaaat atttggagcg aagacaacgc tgattcaggt
     6301 caaatcgatc tcgagatatc gctagttcta gattgaggca tcaaataaaa cgaaaggctc
     6361 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     6421 ggacaaatcc gccgctctag ctaagcagaa ggccatcctg acggatggcc tttttgcgtt
     6481 tctacaaact cttgttgggg atccgcctgc agccaagctt atcgattcta gactcgagag
     6541 ctctcgagtc tagaatcgat acgattttga agtggcaaca gataaaaaaa agcagtttaa
     6601 aattgttgct gaacttttaa aacaagcaaa tacaatcatt gtcgcaacag atagcgacag
     6661 agaaggcgaa aacattgcct ggtcgatcat tcataaagca aatgcctttt ctaaagataa
     6721 aacgtataaa agactatgga tcaatagttt agaaaaagat gtgatccgta gcggttttca
     6781 aaatttgcaa ccaggaatga attactatcc cttttatcaa gaagcgcaca aaaagaaaaa
     6841 cgaaatgata caccaatcag tgcaaaaaaa gatataatgg gagataagac ggttcgtgtt
     6901 cgtgctgact tgcaccatat cataaaaatc gaaacagcaa agaatggcgg aaacgtaaaa
     6961 gaagttatgg aaataagact tagaagcaaa cttaagagtg tgttgatagt gcagtatctt
     7021 aaaattttgt ataataggaa ttgaagttaa attagatgct aaaaatttgt aattaagaag
     7081 gagtgattac atgaacaaaa atataaaata ttctcaaaac tttttaacga gtgaaaaagt
     7141 actcaaccaa ataataaaac aattgaattt aaaagaaacc gataccgttt acgaaattgg
     7201 aacaggtaaa gggcatttaa cgacgaaact ggctaaaata agtaaacagg taacgtctat
     7261 tgaattagac agtcatctat tcaacttatc gtcagaaaaa ttaaaactga atactcgtgt
     7321 cactttaatt caccaagata ttctacagtt tcaattccct aacaaacaga ggtataaaat
     7381 tgttgggagt attccttacc atttaagcac acaaattatt aaaaaagtgg tttttgaaag
     7441 ccatgcgtct gacatctatc tgattgttga agaaggattc tacaagcgta ccttggatat
     7501 tcaccgaaca ctagggttgc tcttgcacac tcaagtctcg attcagcaat tgcttaagct
     7561 gccagcggaa tgctttcatc ctaaaccaaa agtaaacagt gtcttaataa aacttacccg
     7621 ccataccaca gatgttccag ataaatattg gaagctatat acgtactttg tttcaaaatg
     7681 ggtcaatcga gaatatcgtc aactgtttac taaaaatcag tttcatcaag caatgaaaca
     7741 cgccaaagta aacaatttaa gtaccgttac ttatgagcaa gtattgtcta tttttaatag
     7801 ttatctatta tttaacggga ggaaataatt ctatgagtcg cttttgtaaa tttggaaagt
     7861 tacacgttac taaagggaat gtagataaat tattaggtat actactgaca gcttccaagg
     7921 agctaaagag gtccctagcg cttagaatcg ctttaggaaa cacgatccag tccaataatc
     7981 gtcgataaaa acttttgaaa aaggttggtg aaattaccta cttttggaat aatcacaaat
     8041 cacaagtgat taatcacaaa tcacaagtga ttaatcactt gtttattaag atattaaaag
     8101 ctataattta aataaagcgt gaattttatt acacaaaaag aggggggaga aacttggaac
     8161 tagcatttag agaaagctta aaaaagatga gaggtaccaa atcaaaagaa aaattctccc
     8221 aagaattaga aatgagtaga tcaaattatt cacgaataga atcaggaaaa tcagatccaa
     8281 ccataaaaac actagaacaa attgcaaagt taact
//
