LOCUS       V019319                 7244 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019319
VERSION     V019319
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"
     CDS             671..1924
                     /label="AVPR1A(NM_000706)"
                     /note="AVPR1A(NM_000706)"
                     /gene="AVPR1A"
     CDS             1931..1996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2020..2608
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2674..2722
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2924..3352
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3366..3695
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3762..4556
                     /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    4732..4853
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4902..4918)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4926..4942
                     /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(4950..4980)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4995..5016
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5304..5892)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6063..6923)
                     /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(6924..7028)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atgcgtctct ccgccggtcc cgacgcgggg ccctcgggca actccagccc
      721 atggtggcct ctggccaccg gcgctggcaa cacaagccgg gaggccgaag ccctcgggga
      781 gggcaacggc ccaccgaggg acgtgcgcaa cgaggagctg gccaaactgg agatcgccgt
      841 gctggcggtg actttcgcgg tggccgtgct gggcaacagc agcgtactgc tggctctgca
      901 ccggacgccg cgcaagacgt cccgcatgca cctcttcatc cgacacctca gcctggccga
      961 cctggccgtg gcattcttcc aggtgctgcc gcaaatgtgc tgggacatca cctaccgctt
     1021 ccgcggcccc gactggctgt gccgcgtggt gaagcacctg caggtgttcg gcatgtttgc
     1081 gtcggcctac atgctggtag tcatgacagc cgaccgctac atcgcggtgt gccacccgct
     1141 caagactctg caacagcccg cgcgccgctc gcgcctcatg atcgcggccg cctgggtgct
     1201 gagcttcgtg ctgagcacgc cgcagtactt cgtcttctcc atgatcgagg tgaacaatgt
     1261 caccaaggcc cgcgactgct gggccacctt catccagccc tggggttctc gtgcctacgt
     1321 gacctggatg acgggcggca tctttgtggc gcccgtggtc atcttgggta cctgctacgg
     1381 cttcatctgc tacaacatct ggtgcaacgt ccgcgggaag acggcgtcgc gccagagcaa
     1441 gggtgcagag caagcgggtg tggccttcca aaaggggttc ctgctcgcac cctgtgtcag
     1501 cagcgtgaag tccatttccc gggccaagat ccgcacggtg aagatgactt ttgtgatcgt
     1561 gacggcttac atcgtctgct gggcgccttt cttcatcatc cagatgtggt ctgtctggga
     1621 tcccatgtcc gtctggaccg aatcggaaaa ccctaccatc accatcactg cattactggg
     1681 ttccttgaat agctgctgta atccctggat atacatgttt tttagtggcc atctccttca
     1741 agactgtgtt caaagcttcc catgctgcca aaacatgaag gaaaaattca acaaagaaga
     1801 tactgacagt atgagcagaa gacagacttt ttattctaac aatcgaagcc caacaaacag
     1861 tacgggtatg tggaaggact cgcctaaatc ttccaagtcc atcaaattca ttcctgtttc
     1921 aactctcgag gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa
     1981 ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct ggattacaaa
     2041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     2341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     2401 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     2461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2581 cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga
     2641 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2701 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     2761 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     2821 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     2881 gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2941 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3001 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3061 cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc
     3121 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3181 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3241 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt
     3301 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3361 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3421 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3481 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3541 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3601 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3661 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3721 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4561 gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     4621 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     4681 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4861 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4921 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4981 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     5041 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     5101 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5161 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5221 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5281 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5341 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5401 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5461 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5521 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5581 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5641 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5701 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5761 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5821 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5881 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5941 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6001 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6061 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6121 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6181 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6241 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6301 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6361 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6421 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6481 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6541 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6601 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6661 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6721 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6781 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6841 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6901 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6961 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7021 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     7081 atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     7141 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     7201 caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
