LOCUS       V032885                 8751 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032885
VERSION     V032885
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /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"
     CDS             1500..3512
                     /label="Hps4(NM_001359853)"
                     /note="Hps4(NM_001359853)"
                     /gene="Hps4"
     misc_feature    3527..4115
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4181..4229
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4431..4859
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4873..5202
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5269..6063
                     /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    6239..6360
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6409..6425)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6433..6449
                     /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(6457..6487)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6502..6523
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6811..7399)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7570..8430)
                     /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(8431..8535)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tggccaccac cactccccca gagacaaagt cagcagcgtg gtggaactac ttcttcctgt
     1561 atgacggttc caaggtaaag ggagaaggag acccaaccag agctggcatc tgctacttct
     1621 atccgcccca gaccctgtta gaccagcagg aactgctctg tggccagctt gctggagtag
     1681 tgcgctgcct gtgggacctt tctggcaccc cgcccatgct catccgtatg cggaacctca
     1741 agtttgccat cagggcagat ggggactacc tgtgggctct ggggtgtggt gtggagatct
     1801 ctgatgccag ctgcaggcag tttctggatc agctcattgg attttttcat ttctacatgg
     1861 gccctgtctc cctggcctac aagagccatc ctcaggagga gctgagcttg cagtgggata
     1921 cctccatcac acaagtcctc aggagcacca gcgaatcgca caggatcttc aatgccctgt
     1981 ggaacctgga ccgcaccaaa gtggagcccc tgctgctgct gaaggcagct ctcatcttac
     2041 agacctgcca acgctcacct catgtgctag ctggctgcat cctctataaa ggactgattg
     2101 tgaattccca gctcctgcct tccctcactg ctaaggtcct gcttcatcag actgtacctg
     2161 cagaccagag gctgcctgga gcaggagctg ccccacagga aacaggagca gcgttgcccc
     2221 cggatgtgca gatcacctct gtttttctga gtgaggagga agttgccagc ctccacgagt
     2281 tcccagtaga gcatgaaact cgactccagg gaagttcagt tcaataccct ccctgggatc
     2341 aaagcagccc cacccaggca gaagatgcct gggcatcagc tgccatcccg gagcccacgc
     2401 cgcatgatgg agcatgtccg agtggcagcg gggcagatga acgcttgcct aggcttgagc
     2461 aggagtgtgc tgggcccacg ggactgtgca ccactgcctg tggccagggc tccggcctca
     2521 gcagcaggct gcagaaggag ctgtgtctct cccgggagga actggactcg tctgaaatgc
     2581 acgtttctga ggctcaggaa gccttcccac ctttgcctgc cctgggtgat ctggagactc
     2641 tccacagcag ccacagtgcc cccactctac ctgaagacac agccatttgc agctgcctgc
     2701 acccttgccc tcttgagagg ctccctgaga gtggaaggct aggacagctc gcagacctgc
     2761 ccctcaccaa tggccaaacc caggttcctg gcacagaccc tctccccagc agcatgcctg
     2821 tggcacttcc tccccagcat cctgtgggtg tagagcccag tgtggagcca tacgggaatg
     2881 gagcccagga gagccactca gccctgccac gcagttcccg ctcacctgac tctccgggcc
     2941 ccagcccctc tgcagacaga actggcttca agccatcccc atctggacgc cacgcagggc
     3001 tcgtgcccat gaacctctac acccacagtg tgaacggact ggtgctgtcc ctgctggccg
     3061 aagagactct tctcagcgac actgcagcca tcgaggaggt gtaccacagc agcctggcgt
     3121 cgctgaatgg gctggaagtc cacctgaagg agacgctgcc cagagacgag gctagcctca
     3181 ccagcagcac atacaacttc cttcattatg accgcatcca gagcgtgctc tcagccaacc
     3241 tgcccctagt gactgctccc caggaccgcc ggtttctgca ggccgtcaac ctcatgcact
     3301 ccgactttgc cctgctgcct atgctgtacg agatgaccat caggaacgcc tccacagccg
     3361 tgtacgcctg cagcagccca gcccaggaga cctacttcca gcagctggca cccactgccc
     3421 gcagctctgg cttcccaaac cctcaggatt gcgccttcag cctcgcaggc aaagccaagc
     3481 agaagctgct gaagcacggg gtaaacctgc tgtgagatat ctcgacaatc aacctctgga
     3541 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3601 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3661 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3721 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3781 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3841 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3901 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3961 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4021 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4081 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     4141 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4201 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4261 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4321 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4381 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     4441 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4501 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4561 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     4621 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4681 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4741 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4801 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4861 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4921 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4981 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5041 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5101 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5161 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5221 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5281 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5341 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5401 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5461 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5521 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5581 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5641 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5701 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5761 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5821 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5881 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5941 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6001 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6061 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6121 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6181 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6241 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6301 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6361 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6481 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     7021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7381 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7501 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7561 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7621 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7681 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7741 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7801 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7861 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7921 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7981 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8041 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8101 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8161 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8221 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8281 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8341 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8401 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8461 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8521 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8581 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     8641 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     8701 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
