LOCUS       V023911                 8117 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023911
VERSION     V023911
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             779..2797
                     /label="Dpysl2(NM_001378767)"
                     /note="Dpysl2(NM_001378767)"
                     /gene="Dpysl2"
     CDS             2804..2869
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2893..3481
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3547..3595
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3797..4225
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4239..4568
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4635..5429
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5605..5726
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5775..5791)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5799..5815
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5823..5853)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5868..5889
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6177..6765)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6936..7796)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7797..7901)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 ggccgagaga aagcaatctg gaaaggcagc agaggacgaa gaggtccctg ccttttttaa
      841 aaacctgggc tccggcagtc ccaagccgcg gcagaaattc tgcggcatgt tctgccctgt
      901 ggaagggtcc tcggagaaca agaccatcga cttcgactcg ctttcagtgg gccggggctc
      961 ggggcaagtg gtggctcagc agcgcgatgt cgcccacttg ggccccgacc cgcaccggca
     1021 gagccggcgt gccagtgggg agccatctgt tgaatcgagc cggaaggtgg agatccggcg
     1081 ggcctcgggc aaggaagccc tgcagaacat ctatgatcag agcgatcgtc ttctgatcaa
     1141 aggtggcaag attgtgaatg atgaccagtc cttctatgca gacatataca tggaagatgg
     1201 gttgatcaag caaataggag aaaacctgat tgtaccagga ggggtgaaga ccatcgaagc
     1261 ccactccaga atggtgattc ccggagggat tgacgtgcat actcgcttcc agatgcctga
     1321 ccagggaatg acatccgctg atgacttctt ccagggaacc aaggcggccc tggccggggg
     1381 aaccaccatg atcattgacc atgttgttcc tgagcccggg acgagcctat tggctgcctt
     1441 tgatcagtgg agggagtggg ctgacagcaa gtcctgctgt gactattcgc tgcacgtgga
     1501 catcactgag tggcacaagg gcatccagga ggagatggaa gctctggtga aggaccacgg
     1561 ggtaaactcc ttcctcgtgt acatggcttt caaagatcga ttccagctga cggattccca
     1621 gatctatgaa gtgctgagcg tgatccggga tatcggtgcc atagctcaag tccacgcaga
     1681 gaatggtgac atcattgctg aggaacagca gaggatcctg gatctgggca tcacgggccc
     1741 cgagggacac gtgttgagcc ggccagagga ggtcgaggct gaagctgtga accggtccat
     1801 cactattgcc aaccagacca actgccctct gtatgtcacc aaagtgatga gcaagagtgc
     1861 ggctgaagtc atcgctcagg cacggaagaa gggaactgtg gtgtatggtg agcccatcac
     1921 ggccagcctg gggactgatg gctctcatta ctggagcaag aactgggcca aggctgcggc
     1981 ctttgtcacc tccccaccct tgagccccga cccaaccact ccagactttc tcaactcgtt
     2041 gctgtcctgt ggagacctcc aagtcactgg cagtgcccac tgcaccttca acactgccca
     2101 gaaggctgtg gggaaggaca acttcacctt gattcccgag ggcaccaacg gcactgagga
     2161 gcggatgtct gtcatttggg ataaagctgt ggtcactggg aagatggatg agaatcagtt
     2221 tgtggctgtg accagcacca acgcagccaa agtcttcaac ctttaccccc ggaaaggtcg
     2281 catctcggtg ggatctgatg ctgacttggt catctgggac cctgacagtg tgaagaccat
     2341 ctctgccaag acacacaaca gtgctcttga gtacaacatc tttgaaggca tggagtgtcg
     2401 cggctcccca ctggtggtca tcagccaggg caagattgtc ctggaggacg gcacacttca
     2461 tgtcactgaa ggctcaggac gctacattcc ccggaagccc ttccctgact ttgtgtacaa
     2521 acgcatcaaa gcaaggagca ggctggctga gctgagaggg gtccctcgtg gcctgtatga
     2581 cggaccggta tgcgaggtgt ctgtgacgcc caagacggtg actccagcct catcagctaa
     2641 gacatcccct gccaagcagc aggcaccacc tgttcggaac ctgcaccagt ctggattcag
     2701 cttgtctggt gctcagattg acgacaacat tccccgccgc accacccagc gcatcgtggc
     2761 accccctggt ggccgtgcca acatcaccag cctgggcctc gaggactaca aagaccatga
     2821 cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc
     2881 cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     2941 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3001 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3061 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3121 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3181 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3241 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     3301 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3361 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3421 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3481 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     3541 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     3601 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     3661 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     3721 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     3781 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3841 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3901 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3961 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4021 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4081 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4141 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     4201 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4501 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4561 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     4621 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4681 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4741 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4801 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4861 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4921 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4981 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5041 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5101 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5161 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5221 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5281 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5341 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5401 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     5461 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5521 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5581 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5701 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5761 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5821 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5881 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5941 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     6001 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     6061 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     6121 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6181 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6241 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6301 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6361 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6421 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6481 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6541 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6601 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6661 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6721 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6781 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6841 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6901 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6961 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7021 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7081 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7141 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7201 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7261 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7321 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7381 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7441 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7501 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7561 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7621 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7681 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7741 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7801 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7861 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7921 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     7981 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8041 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8101 tgaagaatct gcttagg
//
