LOCUS       V022963                 8580 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022963
VERSION     V022963
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             907..1614
                     /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             1636..4146
                     /label="OPA1(NM_001354663)"
                     /note="OPA1(NM_001354663)"
                     /gene="OPA1"
     polyA_signal    4180..4404
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4450..4878
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4892..5221
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5288..6082
                     /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    6256..6377
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6426..6442)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6450..6466
                     /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(6474..6504)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6519..6540
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6828..7413)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7584..8444)
                     /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(8445..8549)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
      961 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     1021 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     1081 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     1141 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     1201 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     1261 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1321 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1381 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1441 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1501 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1561 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggagga
     1621 gggggaagcg gtaccatgat accggacctt agtgaatata aatggattgt gcctgacatt
     1681 gtgtgggaaa ttgatgagta tatcgatttt ggttctccgg aagaaacggc gtttagagca
     1741 acagatcgtg gatctgaaag tgacaagcat tttagaaagg gtctgcttgg tgagctcatt
     1801 ctcttacaac aacaaattca agagcatgaa gaggaagcgc gcagagccgc tggccaatat
     1861 agcacgagct atgcccaaca gaagcgcaag gtgtcagaca aagagaaaat tgaccaactt
     1921 caggaagaac ttctgcacac tcagttgaag tatcagagaa tcttggaacg attagaaaag
     1981 gagaacaaag aattgagaaa attagtattg cagaaagatg acaaaggcat tcatcataga
     2041 aagcttaaga aatctttgat tgacatgtat tctgaagttc ttgatgttct ctctgattat
     2101 gatgccagtt ataatacgca agatcatctg ccacgggttg ttgtggttgg agatcagagt
     2161 gctggaaaga ctagtgtgtt ggaaatgatt gcccaagctc gaatattccc aagaggatct
     2221 ggggagatga tgacacgttc tccagttaag gtgactctga gtgaaggtcc tcaccatgtg
     2281 gccctattta aagatagttc tcgggagttt gatcttacca aagaagaaga tcttgcagca
     2341 ttaagacatg aaatagaact tcgaatgagg aaaaatgtga aagaaggctg taccgttagc
     2401 cctgagacca tatccttaaa tgtaaaaggc cctggactac agaggatggt gcttgttgac
     2461 ttaccaggtg tgattaatac tgtgacatca ggcatggctc ctgacacaaa ggaaactatt
     2521 ttcagtatca gcaaagctta catgcagaat cctaatgcca tcatactgtg tattcaagat
     2581 ggatctgtgg atgctgaacg cagtattgtt acagacttgg tcagtcaaat ggaccctcat
     2641 ggaaggagaa ccatattcgt tttgaccaaa gtagacctgg cagagaaaaa tgtagccagt
     2701 ccaagcagga ttcagcagat aattgaagga aagctcttcc caatgaaagc tttaggttat
     2761 tttgctgttg taacaggaaa agggaacagc tctgaaagca ttgaagctat aagagaatat
     2821 gaagaagagt tttttcagaa ttcaaagctc ctaaagacaa gcatgctaaa ggcacaccaa
     2881 gtgactacaa gaaatttaag ccttgcagta tcagactgct tttggaaaat ggtacgagag
     2941 tctgttgaac aacaggctga tagtttcaaa gcaacacgtt ttaaccttga aactgaatgg
     3001 aagaataact atcctcgcct gcgggaactt gaccggaatg aactatttga aaaagctaaa
     3061 aatgaaatcc ttgatgaagt tatcagtctg agccaggtta caccaaaaca ttgggaggaa
     3121 atccttcaac aatctttgtg ggaaagagta tcaactcatg tgattgaaaa catctacctt
     3181 ccagctgcgc agaccatgaa ttcaggaact tttaacacca cagtggatat caagcttaaa
     3241 cagtggactg ataaacaact tcctaataaa gcagtagagg ttgcttggga gaccctacaa
     3301 gaagaatttt cccgctttat gacagaaccg aaagggaaag agcatgatga catatttgat
     3361 aaacttaaag aggctgttaa ggaagaaagt attaaacgac acaagtggaa tgactttgcg
     3421 gaggacagct tgagggttat tcaacacaat gctttggaag accgatccat atctgataaa
     3481 cagcaatggg atgcagctat ttattttatg gaagaggctc tgcaggctcg tctcaaggat
     3541 actgaaaatg caattgaaaa catggtgggt ccagactgga aaaagaggtg gttatactgg
     3601 aagaatcgga cccaagaaca gtgtgttcac aatgaaacca agaatgaatt ggagaagatg
     3661 ttgaaatgta atgaggagca cccagcttat cttgcaagtg atgaaataac cacagtccgg
     3721 aagaaccttg aatcccgagg agtagaagta gatccaagct tgattaagga tacttggcat
     3781 caagtttata gaagacattt tttaaaaaca gctctaaacc attgtaacct ttgtcgaaga
     3841 ggtttttatt actaccaaag gcattttgta gattctgagt tggaatgcaa tgatgtggtc
     3901 ttgttttggc gtatacagcg catgcttgct atcaccgcaa atactttaag gcaacaactt
     3961 acaaatactg aagttaggcg attagagaaa aatgttaaag aggtattgga agattttgct
     4021 gaagatggtg agaagaagat taaattgctt actggtaaac gcgttcaact ggcggaagac
     4081 ctcaagaaag ttagagaaat tcaagaaaaa cttgatgctt tcattgaagc tcttcatcag
     4141 gagaaataac ttaagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     4201 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     4261 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     4321 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     4381 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     4441 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4501 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4741 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4801 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4861 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4921 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4981 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5041 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5101 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5161 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5221 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     5281 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     5341 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     5401 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     5461 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5521 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5581 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5641 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5701 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5761 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5821 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5881 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5941 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     6001 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     6061 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     6121 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6181 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6241 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6361 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6541 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7021 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7261 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt tgtttgcaag
     7381 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7441 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7501 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7561 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7621 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7681 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7741 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7801 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7861 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7921 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7981 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8041 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8101 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8161 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8221 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8281 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8341 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8401 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8461 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8521 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
