LOCUS       V022505                 6956 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022505
VERSION     V022505
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..1615
                     /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             1622..2470
                     /label="VDAC1(NM_001401008)"
                     /note="VDAC1(NM_001401008)"
                     /gene="VDAC1"
     polyA_signal    2523..2747
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2793..3221
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3235..3564
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3631..4425
                     /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    4599..4720
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4769..4785)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4793..4809
                     /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(4817..4847)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4862..4883
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5171..5756)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5927..6787)
                     /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(6788..6892)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      961 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1021 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     1081 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1141 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     1201 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1261 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1321 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1381 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1441 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1501 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1561 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggaatt
     1621 catggctgtg ccacccacgt atgccgatct tggcaaatct gccagggatg tcttcaccaa
     1681 gggctatgga tttggcttaa taaagcttga tttgaaaaca aaatctgaga atggattgga
     1741 atttacaagc tcaggctcag ccaacactga gaccaccaaa gtgacgggca gtctggaaac
     1801 caagtacaga tggactgagt acggcctgac gtttacagag aaatggaata ccgacaatac
     1861 actaggcacc gagattactg tggaagatca gcttgcacgt ggactgaagc tgaccttcga
     1921 ttcatccttc tcacctaaca ctgggaaaaa aaatgctaaa atcaagacag ggtacaagcg
     1981 ggagcacatt aacctgggct gcgacatgga tttcgacatt gctgggcctt ccatccgggg
     2041 tgctctggtg ctaggttacg agggctggct ggccggctac cagatgaatt ttgagactgc
     2101 aaaatcccga gtgacccaga gcaactttgc agttggctac aagactgatg aattccagct
     2161 tcacactaat gtgaatgacg ggacagagtt tggcggctcc atttaccaga aagtgaacaa
     2221 gaagttggag accgctgtca atcttgcctg gacagcagga aacagtaaca cgcgcttcgg
     2281 aatagcagcc aagtatcaga ttgaccctga cgcctgcttc tcggctaaag tgaacaactc
     2341 cagcctgata ggtttaggat acactcagac tctaaagcca ggtattaaac tgacactgtc
     2401 agctcttctg gatggcaaga acgtcaatgc tggtggccac aagcttggtc taggactgga
     2461 atttcaagca accggttagc tcgagtctag agggcccgtt taaacccgct gatcagcctc
     2521 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2581 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2641 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2701 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2761 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2821 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2881 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2941 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3001 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3061 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3121 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3181 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     3241 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3301 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3361 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3421 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3481 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     3541 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     3601 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     3661 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     3721 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     3781 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     3841 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     3901 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3961 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     4021 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     4081 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     4141 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     4201 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     4261 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     4321 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     4381 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     4441 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     4501 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4561 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4621 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4681 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     4741 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     4801 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     4861 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     4921 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4981 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5041 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5101 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     5161 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     5221 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5281 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5341 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5401 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5461 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5521 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5581 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5641 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5701 gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5761 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5821 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5881 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5941 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6001 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6061 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6121 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6181 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6241 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6301 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6361 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6421 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6481 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6541 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6601 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6661 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6721 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6781 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6841 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6901 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggt
//
