LOCUS       V033314                 9618 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033314
VERSION     V033314
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..4379
                     /label="Trappc9(NM_029640)"
                     /note="Trappc9(NM_029640)"
                     /gene="Trappc9"
     misc_feature    4394..4982
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5048..5096
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5298..5726
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5740..6069
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6136..6930
                     /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    7106..7227
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7276..7292)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7300..7316
                     /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(7324..7354)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7369..7390
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7678..8266)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8437..9297)
                     /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(9298..9402)
                     /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 tgttgttggg gctgaaaaga cactacaaga agcggtgcca aggacgcatg cggaaacacg
     1561 ttggggactt gtgcctccag gctgggatgc tgcaggacgc cctggtacac taccacatgt
     1621 ccgtggagct cctgcgctca gtgaatgact tcctgtggtt aggggctgcc ctggaggggt
     1681 tgtgctcggc ttcagtcatt tatcattatc ccggaggaac tggtggcaag actggggctc
     1741 ggagactcca aggcagctca cttccctcag aagcagccaa cagacaccgg ccaggggcgc
     1801 aagaagttct cattgaccca ggtgctctca ccaccaacgg cataaatcct gacaccagca
     1861 ctgagatcgg aagggccaag aattgcctca gccccgagga catcattgac aaatacaaag
     1921 aggccatctc ctattacagc aagtacaaga acgccggagt gattgagctg gaagcctgtg
     1981 tcaaggccgt gcgcgtccta gcgattcaga agcgtggcat ggaggcttcg gagtttcttc
     2041 agaatgctgt gtacatcaat ctccggcagc tttcggaaga agagaaaatc cagcgctaca
     2101 gcatcctgtc tgagctctac gagctgattg gcttccaccg caagtcagca ttcttcaagc
     2161 gagtggctgc catgcagtgt gtggcgccaa gcattgcaga acctggctgg agggcctgtt
     2221 acaagctcct tttggagacg ctgcctggct acagtctgtc gctagatccc aaggacttca
     2281 gcaaaggcac acaccgaggc tgggctgccg ttcagatgcg tctgctgcat gagctggtct
     2341 atgcctcccg caggatgggg aaccccgcac tgtccgtgcg gcacctctcc ttcctcttgc
     2401 agaccatgct ggacttcctg tctgaccagg aaaagaaaga tgtgactcaa agcctggaga
     2461 actacactgc caagtgtccc gggacgatgg agcccatcac cctcccagat ggtctcacac
     2521 tgcctcccgt gccgtttacc aagttgccca ttgtcaggtg cgtgaagctg ctgagtctcc
     2581 ccaccagcct gcgaccccac aaggtgaaaa gcttgctggg ccagagcatg tccaccaaga
     2641 gccccttcat ctattcgcca atcatcgcac ataaccgtgg agaagaacgc aacaaaaaaa
     2701 tagatttcca gtgggttcaa ggagatgtgt gtgaagttca gctgatggtg tacaacccaa
     2761 tgccgtttga gctccgcgtg gaaaacatgg ggcttctcac cagcggagtg gagtttgagt
     2821 ctcttcctgc agcgctttcc ctccctgcgg aatctggttt gtacccggtg acgctggtcg
     2881 gagtcccaca gactacagga atgatcactg tgaatggtta ccataccaca gtctttgggg
     2941 tcttcagcga ctgccttctg gacaaccttc cgggactgaa gacgggcggc tccaccgtgg
     3001 aagtcatccc tgccttgcca agactgcaga tcagcacatc gctgcccagg tccgcacgct
     3061 cactgcagcc ttctgccggc gatgagatcg ccaccaacgt atctgttcag ctgtacaacg
     3121 gagaaaccca gcagctggct gtcaccttgg agaacatcgg actggagcct ctggagcagc
     3181 tggaggtcac ctcaaagctt ctcaccacca aggaaaagtt gtatggcgac ttcttgagct
     3241 ggaaactgga agagacgctg gcccagtttc ccctgcagcc cgggaaggtg gctactttca
     3301 ccatcaacat caaggcaaag ctggatttct cttgccagga gaatcttctc caggacctga
     3361 gtgatgatgg aatcagcgtg agcggctttc ccctgtccag tcctttccgc caggtggtgc
     3421 ggccccgagt ggagagcaga cccacaaacc cttctgaggg cagcaagacg ggggaccttg
     3481 gccacgtcaa gactctggaa gctgtcctga acttcaagta ctctggaggc ccaggccacg
     3541 tggaagggta ttacaggaac ctctccctgg gactacacgt ggaagtcgag ccttctgtgt
     3601 tttttacccg agtcagcaca cttccagcaa ccagcacccg gcagtgccac ttgctccttg
     3661 atgtcttcaa ctccacagag cacgagctga cagtctgcgc caggaacaac tctgaacttg
     3721 tcctgcatgc cagtgagtgc cagcgaatgg ctattcaagt ggacaagttc aactttgaaa
     3781 gtgtcccaga atcccccgga gagaagggac actttgcaaa tctgaagcag ctggaagaag
     3841 agagacagga ggcacgaggc ctggagatca gcagcaagct ggacatccgc tggagaattc
     3901 cttctctgaa gcgcagtggt gaggcgagcg tggagggcct cctgaaccag ctcatcctgg
     3961 aacacctgca gctggcacct ctgcagtggg atgtgctagt ggatgggcag ccatgtgact
     4021 gtgaggtggc agctgcctgc caggtcggtg accccgtgcg cttggaggtg cggctgacta
     4081 acaggagtcc ccgaagtgtg gggccctttg ccctcaccgt ggtccctttc caggaccacc
     4141 agaacggtgt gcacaactat gacctgcatg atgtcatctc ctttgtgggt tccagtacct
     4201 tctaccttga cacggtacaa ccatcaggcc aatccacctg ccttggggcc ctcctcttcc
     4261 tctacacagg tgacttcttc ctcaacatcc ggttccacga ggactgcaag agcaaggagc
     4321 tgccaccctc ctgggtttgc ttacccagcg tgcatgtgag ggccctcggg gcacaggcct
     4381 gagatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     4441 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     4501 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     4561 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     4621 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     4681 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     4741 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     4801 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     4861 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     4921 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     4981 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5041 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     5101 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5161 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5221 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     5281 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5341 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5401 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     5461 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5521 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5581 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5641 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     5701 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     5761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     5821 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     5881 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     5941 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6001 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6061 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     6121 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     6181 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     6241 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     6301 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     6361 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     6421 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     6481 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     6541 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     6601 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     6661 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     6721 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     6781 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     6841 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     6901 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     6961 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     7021 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7081 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7141 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7201 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7261 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     7501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     7561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     7621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     7681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     7741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     7801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     7861 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     7921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     7981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8101 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     8341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     8401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     8461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     8521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     8581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     8641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     8701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     8761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     8821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     8881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     8941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     9361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     9421 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     9481 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     9541 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     9601 atgaagaatc tgcttagg
//
