LOCUS       V042437                 8064 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042437
VERSION     V042437
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             776..1990
                     /label="SAG(NM_000541)"
                     /note="SAG(NM_000541)"
                     /gene="SAG"
     CDS             2006..2062
                     /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             2063..2773
                     /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    2840..3428
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3494..3542
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3744..4172
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4186..4515
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4582..5376
                     /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    5552..5673
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5722..5738)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5746..5762
                     /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(5770..5800)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5815..5836
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6124..6712)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6883..7743)
                     /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(7744..7848)
                     /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 ggtaccgagc tcggatcccg ccaccatggc
      781 agccagcggg aagaccagca agtccgaacc gaaccatgtt atcttcaaga agatctcccg
      841 ggacaaatcg gtgaccatct acctggggaa cagagactac atagaccatg tcagccaagt
      901 ccagcctgtg gatggtgtcg tgttggttga tcctgatctt gtgaagggaa agaaagtgta
      961 tgtcactctg acctgcgcct tccgctatgg ccaagaggac attgacgtga tcggcttgac
     1021 cttccgcagg gacctgtact tctcccgggt ccaggtgtat cctcctgtgg gggccgcgag
     1081 cacccccaca aaactgcaag agagcctgct taaaaagctg gggagcaaca cgtacccctt
     1141 tctcctgacg tttcctgact acttgccctg ttcagtgatg ttgcagccag ctccacaaga
     1201 ttcagggaag tcctgtgggg ttgactttga ggtcaaagca ttcgccacag acagcaccga
     1261 tgccgaagag gacaaaatcc ccaagaagag ctccgtgcga ttactgatcc gcaaagtaca
     1321 gcatgcccca cttgagatgg gtccccagcc ccgagctgag gcggcctggc agttcttcat
     1381 gtctgacaag cccctgcacc ttgcggtctc tctcaacaaa gagatctatt tccatgggga
     1441 gcccatccct gtgaccgtga ctgtcaccaa taacacagag aagaccgtga agaagattaa
     1501 agcattcgtg gaacaggtgg ccaatgtggt tctctactcg agtgattatt acgtcaagcc
     1561 cgtggctatg gaggaagcgc aagaaaaagt gccaccaaac agcactttga ccaagacgct
     1621 gacgctgctg cccttgctgg ctaacaatcg agaaaggaga ggcattgccc tggatgggaa
     1681 aatcaagcac gaggacacaa accttgcctc cagcaccatc attaaggagg gcatagaccg
     1741 gaccgtcctg ggaatcctgg tgtcttacca gatcaaggtg aagctcacag tgtcaggctt
     1801 tctgggagag ctcacctcca gtgaagtcgc cactgaggtc ccattccgcc tcatgcaccc
     1861 tcagcctgag gacccagcta aggaaagtta tcaggatgca aatttagttt ttgaggagtt
     1921 tgctcgccat aatctgaaag atgcaggaga agctgaggag gggaagagag acaagaatga
     1981 cgttgatgag gaattcggaa gcggagctac taacttcagc ctgctgaagc aggctggcga
     2041 cgtggaggag aaccctggac ctatggtgag caagggcgag gaggataaca tggccatcat
     2101 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     2161 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2221 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     2281 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     2341 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     2401 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     2461 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     2521 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     2581 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     2641 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     2701 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     2761 gctgtacaag taactcgagt ctagagggcc ccttaagggt tcgatcccta ccggttagta
     2821 atgagtttga tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2881 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2941 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3001 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3061 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3121 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3181 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     3241 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3301 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3361 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3421 ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga
     3481 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     3541 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     3601 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     3661 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag
     3721 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3781 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3841 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3901 catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3961 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4021 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4081 ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa
     4141 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4261 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4321 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4441 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4501 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4561 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4621 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4681 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4741 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4801 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4861 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4921 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4981 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5041 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5101 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5161 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5221 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5281 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5341 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat
     5401 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5461 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5521 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5581 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5641 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5701 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5761 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5821 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5881 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5941 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6001 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6061 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6121 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6181 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6241 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6301 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6361 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6421 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6481 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6541 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6601 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6661 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6721 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6781 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6841 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6901 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6961 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7021 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7081 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7141 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7201 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7261 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7321 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7381 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7441 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7501 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7561 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7621 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7681 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7741 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7801 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7861 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc
     7921 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     7981 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     8041 aattgcatga agaatctgct tagg
//
