LOCUS       V027510                 9731 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027510
VERSION     V027510
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             775..3657
                     /label="GABBR1(NM_001470)"
                     /note="GABBR1(NM_001470)"
                     /gene="GABBR1"
     CDS             3673..3729
                     /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             3730..4440
                     /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    4507..5095
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5161..5209
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5411..5839
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5853..6182
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6249..7043
                     /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    7219..7340
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7389..7405)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7413..7429
                     /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(7437..7467)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7482..7503
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7791..8379)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8550..9410)
                     /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(9411..9515)
                     /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 ggtaccgagc tcggatccgc caccatgttg
      781 ctgctgctgc tactggcgcc actcttcctc cgccccccgg gcgcgggcgg ggcgcagacc
      841 cccaacgcca cctcagaagg ttgccagatc atacacccgc cctgggaagg gggcatcagg
      901 taccggggcc tgactcggga ccaggtgaag gctatcaact tcctgccagt ggactatgag
      961 attgagtatg tgtgccgggg ggagcgcgag gtggtggggc ccaaggtccg caagtgcctg
     1021 gccaacggct cctggacaga tatggacaca cccagccgct gtgtccgaat ctgctccaag
     1081 tcttatttga ccctggaaaa tgggaaggtt ttcctgacgg gtggggacct cccagctctg
     1141 gacggagccc gggtggattt ccggtgtgac cccgacttcc atctggtggg cagctcccgg
     1201 agcatctgta gtcagggcca gtggagcacc cccaagcccc actgccaggt gaatcgaacg
     1261 ccacactcag aacggcgcgc agtgtacatc ggggcactgt ttcccatgag cgggggctgg
     1321 ccagggggcc aggcctgcca gcccgcggtg gagatggcgc tggaggacgt gaatagccgc
     1381 agggacatcc tgccggacta tgagctcaag ctcatccacc acgacagcaa gtgtgatcca
     1441 ggccaagcca ccaagtacct atatgagctg ctctacaacg accctatcaa gatcatcctt
     1501 atgcctggct gcagctctgt ctccacgctg gtggctgagg ctgctaggat gtggaacctc
     1561 attgtgcttt cctatggctc cagctcacca gccctgtcaa accggcagcg tttccccact
     1621 ttcttccgaa cgcacccatc agccacactc cacaacccta cccgcgtgaa actctttgaa
     1681 aagtggggct ggaagaagat tgctaccatc cagcagacca ctgaggtctt cacttcgact
     1741 ctggacgacc tggaggaacg agtgaaggag gctggaattg agattacttt ccgccagagt
     1801 ttcttctcag atccagctgt gcccgtcaaa aacctgaagc gccaggatgc ccgaatcatc
     1861 gtgggacttt tctatgagac tgaagcccgg aaagtttttt gtgaggtgta caaggagcgt
     1921 ctctttggga agaagtacgt ctggttcctc attgggtggt atgctgacaa ttggttcaag
     1981 atctacgacc cttctatcaa ctgcacagtg gatgagatga ctgaggcggt ggagggccac
     2041 atcacaactg agattgtcat gctgaatcct gccaataccc gcagcatttc caacatgaca
     2101 tcccaggaat ttgtggagaa actaaccaag cgactgaaaa gacaccctga ggagacagga
     2161 ggcttccagg aggcaccgct ggcctatgat gccatctggg ccttggcact ggccctgaac
     2221 aagacatctg gaggaggcgg ccgttctggt gtgcgcctgg aggacttcaa ctacaacaac
     2281 cagaccatta ccgaccaaat ctaccgggca atgaactctt cgtcctttga gggtgtctct
     2341 ggccatgtgg tgtttgatgc cagcggctct cggatggcat ggacgcttat cgagcagctt
     2401 cagggtggca gctacaagaa gattggctac tatgacagca ccaaggatga tctttcctgg
     2461 tccaaaacag ataaatggat tggagggtcc cccccagctg accagaccct ggtcatcaag
     2521 acattccgct tcctgtcaca gaaactcttt atctccgtct cagttctctc cagcctgggc
     2581 attgtcctag ctgttgtctg tctgtccttt aacatctaca actcacatgt ccgttatatc
     2641 cagaactcac agcccaacct gaacaacctg actgctgtgg gctgctcact ggctttagct
     2701 gctgtcttcc ccctggggct cgatggttac cacattggga ggaaccagtt tcctttcgtc
     2761 tgccaggccc gcctctggct cctgggcctg ggctttagtc tgggctacgg ttccatgttc
     2821 accaagattt ggtgggtcca cacggtcttc acaaagaagg aagaaaagaa ggagtggagg
     2881 aagactctgg aaccctggaa gctgtatgcc acagtgggcc tgctggtggg catggatgtc
     2941 ctcactctcg ccatctggca gatcgtggac cctctgcacc ggaccattga gacatttgcc
     3001 aaggaggaac ctaaggaaga tattgacgtc tctattctgc cccagctgga gcattgcagc
     3061 tccaggaaga tgaatacatg gcttggcatt ttctatggtt acaaggggct gctgctgctg
     3121 ctgggaatct tccttgctta tgagaccaag agtgtgtcca ctgagaagat caatgatcac
     3181 cgggctgtgg gcatggctat ctacaatgtg gcagtcctgt gcctcatcac tgctcctgtc
     3241 accatgattc tgtccagcca gcaggatgca gcctttgcct ttgcctctct tgccatagtt
     3301 ttctcctcct atatcactct tgttgtgctc tttgtgccca agatgcgcag gctgatcacc
     3361 cgaggggaat ggcagtcgga ggcgcaggac accatgaaga cagggtcatc gaccaacaac
     3421 aacgaggagg agaagtcccg gctgttggag aaggagaacc gtgaactgga aaagatcatt
     3481 gctgagaaag aggagcgtgt ctctgaactg cgccatcaac tccagtctcg gcagcagctc
     3541 cgctcccggc gccacccacc gacaccccca gaaccctctg ggggcctgcc caggggaccc
     3601 cctgagcccc ccgaccggct tagctgtgat gggagtcgag tgcatttgct ttataaggaa
     3661 ttcggaagcg gagctactaa cttcagcctg ctgaagcagg ctggcgacgt ggaggagaac
     3721 cctggaccta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     3781 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     3841 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     3901 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     3961 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     4021 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     4081 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac
     4141 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc
     4201 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     4261 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     4321 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     4381 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     4441 ctcgagtcta gagggcccct taagggttcg atccctaccg gttagtaatg agtttgatat
     4501 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4561 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4621 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4681 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4741 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4801 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4861 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4921 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4981 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     5041 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     5101 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5161 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     5221 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5281 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5341 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     5401 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     5461 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     5521 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     5581 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     5641 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5701 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     5761 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     5821 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     5881 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     5941 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     6001 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     6061 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     6121 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     6181 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     6241 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     6301 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6361 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6421 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6481 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6541 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6601 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     6661 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     6721 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     6781 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     6841 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     6901 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     6961 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7021 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     7081 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7141 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7201 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7261 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7321 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7381 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7441 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7501 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7561 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7621 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7681 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7741 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7801 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7861 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7921 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7981 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8041 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8101 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8161 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8221 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8281 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8341 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8401 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8461 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8521 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8581 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8641 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8701 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8761 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8821 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8881 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8941 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9001 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9061 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9541 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     9601 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9661 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9721 atctgcttag g
//
