LOCUS       V019297                 7415 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019297
VERSION     V019297
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..2095
                     /label="GABRG2(NM_198904)"
                     /note="GABRG2(NM_198904)"
                     /gene="GABRG2"
     CDS             2102..2167
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2191..2779
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2845..2893
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3095..3523
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3537..3866
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3933..4727
                     /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    4903..5024
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5073..5089)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5097..5113
                     /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(5121..5151)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5166..5187
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5475..6063)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6234..7094)
                     /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(7095..7199)
                     /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 atgagttcgc caaatatatg gagcacagga agctcagtct actcgactcc
      721 tgtattttca cagaaaatga cggtgtggat tctgctcctg ctgtcgctct accctggctt
      781 cactagccag aaatctgatg atgactatga agattatgct tctaacaaaa catgggtctt
      841 gactccaaaa gttcctgagg gtgatgtcac tgtcatctta aacaacctgc tggaaggata
      901 tgacaataaa cttcggcctg atataggagt gaagccaacg ttaattcaca cagacatgta
      961 tgtgaatagc attggtccag tgaacgctat caatatggaa tacactattg atatattttt
     1021 tgcgcaaacg tggtatgaca gacgtttgaa atttaacagc accattaaag tcctccgatt
     1081 gaacagcaac atggtgggga aaatctggat tccagacact ttcttcagaa attccaaaaa
     1141 agctgatgca cactggatca ccacccccaa caggatgctg agaatttgga atgatggtcg
     1201 agtgctctac accctaaggt tgacaattga tgctgagtgc caattacaat tgcacaactt
     1261 tccaatggat gaacactcct gccccttgga gttctcaagt tatggctatc cacgtgaaga
     1321 aattgtttat caatggaagc gaagttctgt tgaagtgggc gacacaagat cctggaggct
     1381 ttatcaattc tcatttgttg gtctaagaaa taccaccgaa gtagtgaaga caacttccgg
     1441 agattatgtg gtcatgtctg tctactttga tctgagcaga agaatgggat actttaccat
     1501 ccagacctat atcccctgca cactcattgt cgtcctatcc tgggtgtctt tctggatcaa
     1561 taaggatgct gttccagcca gaacatcttt aggtatcacc actgtcctga caatgaccac
     1621 cctcagcacc attgcccgga aatcgctccc caaggtctcc tatgtcacag cgatggatct
     1681 ctttgtatct gtttgtttca tctttgtctt ctctgctctg gtggagtatg gcaccttgca
     1741 ttattttgtc agcaaccgga aaccaagcaa ggacaaagat aaaaagaaga aaaaccctct
     1801 tcttcggatg ttttccttca aggcccctac cattgatatc cgcccaagat cagcaaccat
     1861 tcaaatgaat aatgctacac accttcaaga gagagatgaa gagtacggct atgagtgtct
     1921 ggacggcaag gactgtgcca gttttttctg ctgttttgaa gattgtcgaa caggagcttg
     1981 gagacatggg aggatacata tccgcattgc caaaatggac tcctatgctc ggatcttctt
     2041 ccccactgcc ttctgcctgt ttaatctggt ctattgggtc tcctacctct acctgctcga
     2101 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2161 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2221 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2281 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2341 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2401 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2461 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2521 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2581 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2641 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2701 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2761 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     2821 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     2881 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     2941 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3001 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3061 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3121 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3181 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3241 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3301 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3361 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3421 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3481 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3541 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3601 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3721 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3781 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3841 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     3901 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3961 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4021 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4081 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4141 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4201 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4261 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4321 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4381 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4441 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4501 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4561 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4621 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4681 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4741 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4801 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4861 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     4921 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4981 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5041 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5101 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5161 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5221 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5281 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5341 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5401 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5461 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5521 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5581 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5641 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5701 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5761 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5821 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5881 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     5941 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6001 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6061 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6121 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6181 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6241 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6301 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6361 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6421 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6481 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6541 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6601 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6661 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6721 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6781 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6841 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6901 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6961 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7021 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7081 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7141 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7201 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7261 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7321 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7381 gcttgaccga caattgcatg aagaatctgc ttagg
//
