LOCUS       V029165                 7801 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029165
VERSION     V029165
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             706..2460
                     /label="Gad2(NM_008078)"
                     /note="Gad2(NM_008078)"
                     /gene="Gad2"
     CDS             2467..2493
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2497..2523
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2527..2553
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2577..3165
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3231..3279
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3481..3909
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3923..4252
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4319..5113
                     /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    5289..5410
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5459..5475)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5483..5499
                     /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(5507..5537)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5552..5573
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5861..6449)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6620..7480)
                     /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(7481..7585)
                     /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 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gtctcctggc
      721 tccggctttt ggtccttcgg atctgaagat ggctctgcgg atcctgagaa cccgggcaca
      781 gcgagagcgt ggtgccaggt ggcccaaaag ttcacgggcg gcatcggaaa caagctgtgc
      841 gctctgctct atggagactc cgggaagcca gcggagggcg gcgggagcgt gacctcgcgg
      901 gccgccaccg ggaaggtcgc ctgcacctgc gaccaaaaac cctgcaactg ccccaaaggg
      961 gatgtcaact acgcgtttct gcacgcaaca gacctgctgc cagcgtgtga tggagaaagg
     1021 cccactctgg catttctaca agatgttatg aacattttgc ttcagtacgt ggtgaaaagt
     1081 ttcgatagat caactaaagt gatcgatttc cattacccca atgagcttct tcaagagtat
     1141 aattgggaat tggcagacca accgcaaaat ctggaggaaa ttttgacgca ttgccaaaca
     1201 actctaaaat atgcaattaa aacagggcat ccccgatatt ttaatcagct gtctactgga
     1261 ttggatatgg ttggattagc agcagactgg ttgacatcaa cagcaaacac taacatgttc
     1321 acctatgaga tcgcccctgt atttgtgctg ctagaatatg ttacactaaa gaaaatgaga
     1381 gaaatcattg gctggcccgg gggctctggc gatggaatct tttctcctgg tggcgccatc
     1441 tccaacatgt atgccatgct cattgcccgc tataagatgt ttccagaagt caaggagaag
     1501 gggatggcag cagtgcccag gctcatcgcg ttcacatcag agcatagtca cttttctctc
     1561 aagaagggag ctgcagcctt agggattgga acagacagtg tgattctgat taaatgtgac
     1621 gagagaggga aaatgattcc atctgatctt gaaagaagaa tccttgaagt caaacagaaa
     1681 ggatttgttc ctttcctggt gagtgccaca gctggaacca ccgtgtatgg ggcttttgat
     1741 cctctcttgg ctgtagctga catctgcaaa aagtataaga tctggatgca cgtggatgct
     1801 gcttggggtg gagggttact gatgtcccgg aaacacaagt ggaagctgag tggagtagag
     1861 agggccaact ctgtgacatg gaatcctcac aagatgatgg gtgtcccctt gcagtgttca
     1921 gctctcctgg ttagagagga gggactgatg cagagctgca accagatgca tgcttcctac
     1981 ctctttcagc aagataagca ctatgacctg tcctatgaca caggagacaa ggccttgcag
     2041 tgtggacgcc atgtggatgt ctttaaatta tggctcatgt ggagagcaaa ggggactact
     2101 gggtttgagg cacacattga taagtgtttg gagctagcag agtatttata cactatcata
     2161 aaaaaccgag aaggctatga aatggtgttt gatgggaagc ctcaacacac aaatgtctgc
     2221 ttctggtttg tacctcctag tttgcgcact ctggaagaca atgaagagag aatgagccgc
     2281 ctctcaaagg tggcgccagt gattaaagcc agaatgatgg agtatgggac cacaatggtc
     2341 agctaccaac ccttagggga caaggtcaac ttcttccgca tggtcatctc aaaccctgca
     2401 gcaactcacc aagacattga cttccttatt gaagaaatcg aacgcctcgg acaagatttg
     2461 ctcgagtacc catacgacgt acctgattat gcaacttatc cttatgatgt acctgattat
     2521 gctagctatc cttatgatgt cccagactac gcatagtgag ggcccgatat ctcgacaatc
     2581 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2641 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2701 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2761 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2821 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2881 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2941 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     3001 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     3061 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     3121 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag
     3181 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     3241 aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3301 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3361 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3421 tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc
     3481 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3541 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3601 cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta
     3661 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3721 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3781 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3841 aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3901 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3961 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4021 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4081 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4141 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     4201 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     4261 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     4321 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4381 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4441 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4501 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4561 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4621 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4681 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4741 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4801 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4861 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4921 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4981 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     5041 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     5101 cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc
     5161 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     5221 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     5281 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5341 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5401 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct
     6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6301 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6361 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     6961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     7261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     7501 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7561 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     7621 gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc
     7681 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     7741 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     7801 g
//
