LOCUS       Exported                7401 bp DNA     circular SYN 20-JUL-2025
DEFINITION  Exported.
ACCESSION   V016759
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7401)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7401)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7401)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        47..425
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        426..629
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        860..878
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             971..994
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             995..2356
                     /gene="pylS"
                     /label=Pyrrolysine--tRNA ligase
                     /note="Pyrrolysine--tRNA ligase from Methanosarcina mazei 
                     (strain ATCC BAA-159 / DSM 3647 / Goe1 / Go1 / JCM 11833 / 
                     OCM 88). Accession#: Q8PWY1"
     polyA_signal    2391..2615
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2661..3089
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3103..3432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3481..4503
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     polyA_signal    4636..4769
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4806..4822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4830..4846)
                     /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)."
     promoter        complement(4854..4884)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4899..4920)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5970..6827)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6828..6932)
                     /label=AmpR promoter
     tRNA            complement(6953..7021)
                     /product="tRNAPyl"
     promoter        complement(7030..7270)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
        1 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga
       61 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      121 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      181 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      241 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      301 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      361 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      421 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
      481 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      541 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
      601 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
      661 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc
      721 gggaacggtg cattggaacg cggattcccc gtgccaagag tgacgtaagt accgcctata
      781 gagtctatag gcccaccccc ttggcttcgt tagaacgcgg ctacaattaa tacataacct
      841 tatgtatcat acacatacga tttaggtgac actatagaat aacatccact ttgcctttct
      901 ctccacaggt gtccactccc aggtccaact gcacgggcta gcgtttaaac ttaagaagct
      961 tgccaccatg gactacaagg acgacgacga caagatggac aagaagcccc tgaacaccct
     1021 gatcagcgcc acaggactgt ggatgtccag aaccggcacc atccacaaga tcaagcacca
     1081 cgaggtgtcc cggtccaaaa tctacatcga gatggcctgc ggcgatcacc tggtcgtcaa
     1141 caacagcaga agcagccgga cagccagagc cctgcggcac cacaagtaca gaaagacctg
     1201 caagcggtgc agagtgtccg acgaggacct gaacaagttc ctgaccaagg ccaacgagga
     1261 ccagaccagc gtgaaagtga aggtggtgtc cgcccccacc cggaccaaga aagccatgcc
     1321 caagagcgtg gccagagccc ccaagcccct ggaaaacacc gaagccgctc aggcccagcc
     1381 cagcggcagc aagttcagcc ccgccatccc cgtgtctacc caggaaagcg tcagcgtccc
     1441 cgccagcgtg tccaccagca tctctagcat ctcaaccggc gccacagctt ctgccctggt
     1501 caagggcaac accaacccca tcaccagcat gtctgcccct gtgcaggcct ctgccccagc
     1561 cctgaccaag tcccagaccg accggctgga agtgctcctg aaccccaagg acgagatcag
     1621 cctgaacagc ggcaagccct tccgggagct ggaaagcgag ctgctgagcc ggcggaagaa
     1681 ggacctccag caaatctacg ccgaggaacg ggagaactac ctgggcaagc tggaaagaga
     1741 gatcacccgg ttcttcgtgg accggggctt cctggaaatc aagagcccca tcctgatccc
     1801 cctggagtac atcgagcgga tgggcatcga caacgacacc gagctgagca agcagatttt
     1861 ccgggtggac aagaacttct gcctgcggcc catgctggcc cccaacctgg ccaactacct
     1921 gcggaaactg gatcgcgctc tgcccgaccc catcaagatt ttcgagatcg gcccctgcta
     1981 ccggaaagag agcgacggca aagagcacct ggaagagttt acaatgctga acttttgcca
     2041 gatgggcagc ggctgcacca gagagaacct ggaatccatc atcaccgact ttctgaacca
     2101 cctggggatc gacttcaaga tcgtgggcga cagctgcatg gtgttcggcg acaccctgga
     2161 cgtgatgcac ggcgacctgg aactgtctag cgccgtcgtg ggacccatcc ctctggaccg
     2221 ggagtggggc atcgataagc cctggatcgg agccggcttc ggcctggaac ggctgctgaa
     2281 agtcaagcac gactttaaga acatcaagcg ggctgccaga agcgagagct actacaacgg
     2341 catcagcacc aacctgtgat gataaggatc cacccgctga tcagcctcga ctgtgccttc
     2401 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2461 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2521 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2581 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     2641 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2701 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2761 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2821 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2881 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2941 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3001 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3061 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3361 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3421 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     3481 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     3541 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     3601 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     3661 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     3721 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     3781 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     3841 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     3901 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     3961 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     4021 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     4081 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     4141 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     4201 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     4261 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     4321 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     4381 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     4441 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     4501 gaatagcacg tgctacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4561 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4621 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4681 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4741 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     4801 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4861 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4921 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4981 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaaat ggcggaaacc
     6961 ccgggaatct aacccggctg aacggattta gagtccattc gatctacatg atcaggtttc
     7021 cggtgtttcg tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta
     7081 cggtaagcat atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa
     7141 attattactt tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa
     7201 ttctaattat ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa
     7261 ataggccctc ttcctgcccg accttcgcga tgtacgggcc agatatacgc caatattggc
     7321 cattagccat attattcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7381 atacgttgta tctatatcat a
//
