LOCUS       V043189                 9808 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043189
VERSION     V043189
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             819..3482
                     /label="HMGCR(NM_000859)"
                     /note="HMGCR(NM_000859)"
                     /gene="HMGCR"
     CDS             3546..3692
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /transl_table=1
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
                     /label="PDGFRβ TM domain"
     CDS             3750..3806
                     /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             3807..4517
                     /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    4584..5172
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5238..5286
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5488..5916
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5930..6259
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6326..7120
                     /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    7296..7417
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7466..7482)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7490..7506
                     /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(7514..7544)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7559..7580
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7868..8456)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8627..9487)
                     /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(9488..9592)
                     /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 gagccaccat gagggcctgg atcttctttc
      781 tcctttgcct ggccgggagg gccttggcac tgcccgtcat gttgtcaaga ctttttcgaa
      841 tgcatggcct ctttgtggcc tcccatccct gggaagtcat agtggggaca gtgacactga
      901 ccatctgcat gatgtccatg aacatgttta ctggtaacaa taagatctgt ggttggaatt
      961 atgaatgtcc aaagtttgaa gaggatgttt tgagcagtga cattataatt ctgacaataa
     1021 cacgatgcat agccatcctg tatatttact tccagttcca gaatttacgt caacttggat
     1081 caaaatatat tttgggtatt gctggccttt tcacaatttt ctcaagtttt gtattcagta
     1141 cagttgtcat tcacttctta gacaaagaat tgacaggctt gaatgaagct ttgccctttt
     1201 tcctactttt gattgacctt tccagagcaa gcacattagc aaagtttgcc ctcagttcca
     1261 actcacagga tgaagtaagg gaaaatattg ctcgtggaat ggcaatttta ggtcctacgt
     1321 ttaccctcga tgctcttgtt gaatgtcttg tgattggagt tggtaccatg tcaggggtac
     1381 gtcagcttga aattatgtgc tgctttggct gcatgtcagt tcttgccaac tacttcgtgt
     1441 tcatgacttt cttcccagct tgtgtgtcct tggtattaga gctttctcgg gaaagccgcg
     1501 agggtcgtcc aatttggcag ctcagccatt ttgcccgagt tttagaagaa gaagaaaata
     1561 agccgaatcc tgtaactcag agggtcaaga tgattatgtc tctaggcttg gttcttgttc
     1621 atgctcacag tcgctggata gctgatcctt ctcctcaaaa cagtacagca gatacttcta
     1681 aggtttcatt aggactggat gaaaatgtgt ccaagagaat tgaaccaagt gtttccctct
     1741 ggcagtttta tctctctaaa atgatcagca tggatattga acaagttatt accctaagtt
     1801 tagctctcct tctggctgtc aagtacatct tctttgaaca aacagagaca gaatctacac
     1861 tctcattaaa aaaccctatc acatctcctg tagtgacaca aaagaaagtc ccagacaatt
     1921 gttgtagacg tgaacctatg ctggtcagaa ataaccagaa atgtgattca gtagaggaag
     1981 agacagggat aaaccgagaa agaaaagttg aggttataaa acccttagtg gctgaaacag
     2041 ataccccaaa cagagctaca tttgtggttg gtaactcctc cttactcgat acttcatcag
     2101 tactggtgac acaggaacct gaaattgaac ttcccaggga acctcggcct aatgaagaat
     2161 gtctacagat acttgggaat gcagagaaag gtgcaaaatt ccttagtgat gctgagatca
     2221 tccagttagt caatgctaag catatcccag cctacaagtt ggaaactctg atggaaactc
     2281 atgagcgtgg tgtatctatt cgccgacagt tactttccaa gaagctttca gaaccttctt
     2341 ctctccagta cctaccttac agggattata attactcctt ggtgatggga gcttgttgtg
     2401 agaatgttat tggatatatg cccatccctg ttggagtggc aggacccctt tgcttagatg
     2461 aaaaagaatt tcaggttcca atggcaacaa cagaaggttg tcttgtggcc agcaccaata
     2521 gaggctgcag agcaataggt cttggtggag gtgccagcag ccgagtcctt gcagatggga
     2581 tgactcgtgg cccagttgtg cgtcttccac gtgcttgtga ctctgcagaa gtgaaagcct
     2641 ggctcgaaac atctgaaggg ttcgcagtga taaaggaggc atttgacagc actagcagat
     2701 ttgcacgtct acagaaactt catacaagta tagctggacg caacctttat atccgtttcc
     2761 agtccaggtc aggggatgcc atggggatga acatgatttc aaagggtaca gagaaagcac
     2821 tttcaaaact tcacgagtat ttccctgaaa tgcagattct agccgttagt ggtaactatt
     2881 gtactgacaa gaaacctgct gctataaatt ggatagaggg aagaggaaaa tctgttgttt
     2941 gtgaagctgt cattccagcc aaggttgtca gagaagtatt aaagactacc acagaggcta
     3001 tgattgaggt caacattaac aagaatttag tgggctctgc catggctggg agcataggag
     3061 gctacaacgc ccatgcagca aacattgtca ccgccatcta cattgcctgt ggacaggatg
     3121 cagcacagaa tgttggtagt tcaaactgta ttactttaat ggaagcaagt ggtcccacaa
     3181 atgaagattt atatatcagc tgcaccatgc catctataga gataggaacg gtgggtggtg
     3241 ggaccaacct actacctcag caagcctgtt tgcagatgct aggtgttcaa ggagcatgca
     3301 aagataatcc tggggaaaat gcccggcagc ttgcccgaat tgtgtgtggg accgtaatgg
     3361 ctggggaatt gtcacttatg gcagcattgg cagcaggaca tcttgtcaaa agtcacatga
     3421 ttcacaacag gtcgaagatc aatttacaag acctccaagg agcttgcacc aagaagacag
     3481 ccggctccac cagcggcagc ggcaagccag gctccggcga aggcagcacc aaaggcgatc
     3541 tgaatgctgt gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta
     3601 aggtggtggt gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca
     3661 tcctcatcat gctttggcag aagaagccac gtatagggat ccgtctcgag tctagagggc
     3721 ccttcggggg tggaggctct ggaagcggag ctactaactt cagcctgctg aagcaggctg
     3781 gcgacgtgga ggagaaccct ggacctatgg tgagcaaggg cgaggaggat aacatggcca
     3841 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     3901 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     3961 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     4021 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     4081 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     4141 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     4201 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     4261 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     4321 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     4381 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     4441 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     4501 acgagctgta caagtaactc gagtctagag ggccccttaa gggttcgatc cctaccggtt
     4561 agtaatgagt ttgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga
     4621 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     4681 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     4741 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     4801 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     4861 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     4921 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc
     4981 tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     5041 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     5101 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     5161 ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga
     5221 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt
     5281 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     5341 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     5401 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc
     5461 gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     5521 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5581 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5641 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5701 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5761 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5821 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     5881 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     5941 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6001 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6061 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     6121 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6181 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     6241 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     6301 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     6361 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6421 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6481 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     6541 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6601 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6661 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6721 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6781 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6841 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6901 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6961 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     7021 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     7081 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg
     7141 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     7201 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     7261 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     7321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     7441 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7501 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7561 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7621 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7681 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7741 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7801 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     7861 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     7921 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     7981 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8041 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     8101 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8161 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8221 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8281 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     8341 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8401 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8461 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8521 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8581 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8641 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8701 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8761 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     8821 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     8881 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     8941 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9001 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9061 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9121 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9181 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9241 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9301 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9361 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9421 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9481 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9541 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9601 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac
     9661 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     9721 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     9781 cgacaattgc atgaagaatc tgcttagg
//
