LOCUS       V043362                 8665 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043362
VERSION     V043362
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             819..2339
                     /label="NOVA1(NM_002515)"
                     /note="NOVA1(NM_002515)"
                     /gene="NOVA1"
     CDS             2403..2549
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /transl_table=1
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
                     /label="PDGFRβ TM domain"
     CDS             2607..2663
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2664..3374
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    3441..4029
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4095..4143
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4345..4773
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4787..5116
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5183..5977
                     /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    6153..6274
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6323..6339)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6347..6363
                     /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(6371..6401)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6416..6437
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6725..7313)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7484..8344)
                     /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(8345..8449)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt gagccaccat gagggcctgg atcttctttc
      781 tcctttgcct ggccgggagg gccttggcac tgcccgtcat gatggcggca gctcccatcc
      841 agcagaacgg gacccacact ggggttccca tagacctgga cccgccggac tcgcggaaaa
      901 ggccgctgga agccccccct gaagccggca gcaccaagag gaccaatacg ggcgaagacg
      961 gccagtattt tctaaaggtt ctcataccta gttatgctgc tggatctata attgggaagg
     1021 gaggacagac aattgttcag ttgcaaaaag aaactggagc caccatcaag ctgtctaagt
     1081 ccaaagattt ttacccaggt actactgagc gagtgtgctt gatccaggga acggttgaag
     1141 cactgaatgc agttcatgga ttcattgcag aaaaaattcg agaaatgccc caaaatgtgg
     1201 ccaagacaga accagtcagc attctacaac cccagaccac cgttaatcca gatcgcatca
     1261 aacaaacatt gccatcttcc ccaactacca ccaagtcctc tccatctgat cccatgacca
     1321 cctccagagc taatcaggta aagattatag ttcccaacag cacagcaggt ctgataatag
     1381 ggaagggagg tgctactgtg aaggctgtaa tggagcagtc aggggcttgg gtgcagcttt
     1441 cccagaaacc tgatgggatc aacttgcaag agagggttgt cactgtgagt ggagaacctg
     1501 aacaaaaccg aaaagctgtt gaacttatca tccagaagat acaagaggat ccacaaagtg
     1561 gcagctgtct caatatcagt tatgccaatg tgacaggtcc agtggcaaat tccaatccaa
     1621 ccggatctcc ttatgcaaac actgctgaag tgttaccaac tgctgcagca gctgcagggc
     1681 tattaggaca tgctaacctt gctggcgttg cagcctttcc agcagtttta tctggcttca
     1741 caggcaatga cctggtggcc atcacctctg cacttaatac attagccagc tatggatata
     1801 atctcaacac tttaggttta ggtctcagtc aagcagcagc aacaggggct ttggctgcag
     1861 cagctgccag tgccaaccca gcagcagcag cagccaattt attggccacc tatgccagtg
     1921 aagcctcagc cagtggcagc acagctggtg gtacggcggg gacatttgca ttaggtagcc
     1981 tggctgctgc tactgctgca accaatggat attttggagc tgcttctccc ctagctgcca
     2041 gtgccattct aggaacagaa aagtccacag atggatccaa ggatgtagtt gaaatagcag
     2101 tgccagaaaa cttagttggt gcaatacttg gcaaaggagg gaaaacatta gtggaatacc
     2161 aggagttgac tggtgcaagg atacagatct ccaaaaaagg agaattcgta cctggcacaa
     2221 ggaatcggaa ggtaaccatt actggaacac cagctgcaac acaggctgct caatatttaa
     2281 ttacacaaag gatcacatat gagcaaggag ttcgggctgc caatcctcag aaagtgggtg
     2341 gctccaccag cggcagcggc aagccaggct ccggcgaagg cagcaccaaa ggcgatctga
     2401 atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg ccctttaagg
     2461 tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc cttatcatcc
     2521 tcatcatgct ttggcagaag aagccacgta tagggatccg tctcgagtct agagggccct
     2581 tcgggggtgg aggctctgga agcggagcta ctaacttcag cctgctgaag caggctggcg
     2641 acgtggagga gaaccctgga cctatggtga gcaagggcga ggaggataac atggccatca
     2701 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
     2761 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
     2821 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     2881 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     2941 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     3001 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     3061 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     3121 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     3181 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     3241 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     3301 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     3361 agctgtacaa gtaactcgag tctagagggc cccttaaggg ttcgatccct accggttagt
     3421 aatgagtttg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     3481 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3541 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3601 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3661 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3721 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3781 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     3841 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3901 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3961 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4021 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     4081 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     4141 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     4201 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     4261 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     4321 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4381 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     4441 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     4501 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     4561 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     4621 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     4681 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     4741 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     4801 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4861 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4921 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4981 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     5041 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     5101 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     5161 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     5221 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     5281 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     5341 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     5401 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     5461 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     5521 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     5581 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     5641 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     5701 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5761 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5821 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5881 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5941 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     6001 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     6061 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     6121 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6181 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6241 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6301 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6361 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6421 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6481 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6541 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6601 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6661 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6721 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6781 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6841 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6901 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     6961 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7021 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7081 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7141 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7201 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7261 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7321 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7381 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7441 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7501 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7561 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7621 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7681 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7741 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7801 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7861 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7921 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7981 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8041 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8101 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8161 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8221 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8281 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8341 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8401 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8461 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     8521 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     8581 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     8641 caattgcatg aagaatctgc ttagg
//
