LOCUS       V032250                 8927 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032250
VERSION     V032250
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             775..2853
                     /label="TGM3(NM_003245)"
                     /note="TGM3(NM_003245)"
                     /gene="TGM3"
     CDS             2869..2925
                     /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             2926..3636
                     /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    3703..4291
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4357..4405
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4607..5035
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5049..5378
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5445..6239
                     /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    6415..6536
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6585..6601)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6609..6625
                     /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(6633..6663)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6678..6699
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6987..7575)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7746..8606)
                     /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(8607..8711)
                     /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 tcggatccgc caccatggct
      781 gctctaggag tccagagtat caactggcag acggccttca accgacaagc gcatcacaca
      841 gacaagttct ccagccagga gctcatcttg cggagaggcc aaaacttcca ggtcttaatg
      901 atcatgaaca aaggccttgg ctctaacgaa agactggagt tcattgtctc cacagggcct
      961 tacccctcag agtcggccat gacgaaggct gtgtttccac tctccaatgg cagtagtggt
     1021 ggctggagtg cggtgcttca ggccagcaat ggcaatactc tgactatcag catctccagt
     1081 cctgccagcg cacccatagg acggtacaca atggccctcc agatcttctc ccagggcggc
     1141 atctcctctg tgaaacttgg gacgttcata ctgcttttta acccctggct gaatgtggat
     1201 agcgtcttta tgggtaacca cgctgagaga gaagagtatg ttcaggaaga tgccggcatc
     1261 atctttgtgg gaagcacaaa ccgaattggc atgattggct ggaactttgg acagtttgaa
     1321 gaagacattc tcagcatctg cctctcaatc ttggatagga gtctgaattt ccgccgtgac
     1381 gctgctactg atgtggccag cagaaatgac cccaaatacg ttggccgggt gctgagtgcc
     1441 atgatcaata gcaatgatga caatggtgtg cttgctggga attggagcgg cacttacacc
     1501 ggtggccggg acccaaggag ctggaacggc agcgtggaga tcctcaaaaa ttggaaaaaa
     1561 tctggcttca gcccagtccg atatggccag tgctgggtct ttgctgggac cctcaacaca
     1621 gcgctgcggt ctttggggat tccttcccgg gtgatcacca acttcaactc agctcatgac
     1681 acagaccgaa atctcagtgt ggatgtgtac tacgacccca tgggaaaccc cctggacaag
     1741 ggtagtgata gcgtatggaa tttccatgtc tggaatgaag gctggtttgt gaggtctgac
     1801 ctgggcccct cgtacggtgg atggcaggtg ttggatgcta ccccgcagga aagaagccaa
     1861 ggggtgttcc agtgcggccc cgcttcggtc attggtgttc gagagggtga tgtgcagctg
     1921 aacttcgaca tgccctttat cttcgcggag gttaatgccg accgcatcac ctggctgtac
     1981 gacaacacca ctggcaaaca gtggaagaat tccgtgaaca gtcacaccat tggcaggtac
     2041 atcagcacca aggcggtggg cagcaatgct cgcatggacg tcacggacaa gtacaagtac
     2101 ccagaaggct ctgaccagga aagacaagtg ttccaaaagg ctttggggaa acttaaaccc
     2161 aacacgccat ttgccgcgac gtcttcaatg ggtttggaaa cagaggaaca ggagcccagc
     2221 atcatcggga agctgaaggt cgctggcatg ctggcagtag gcaaagaagt caacctggtc
     2281 ctactgctca aaaacctgag cagggatacg aagacagtga cagtgaacat gacagcctgg
     2341 accatcatct acaacggcac gcttgtacat gaagtgtgga aggactctgc cacaatgtcc
     2401 ctggaccctg aggaagaggc agaacatccc ataaagatct cgtacgctca gtatgagaag
     2461 tacctgaagt cagacaacat gatccggatc acagcggtgt gcaaggtccc agatgagtct
     2521 gaggtggtgg tggagcggga catcatcctg gacaacccca ccttgaccct ggaggtgctg
     2581 aacgaggctc gtgtgcggaa gcctgtgaac gtgcagatgc tcttctccaa tccactggat
     2641 gagccggtga gggactgcgt gctgatggtg gagggaagcg gcctgctgtt gggtaacctg
     2701 aagatcgacg tgccgaccct agggcccaag gaggggtccc gggtccgttt tgatatcctg
     2761 ccctcccgga gtggcaccaa gcaactgctc gccgacttct cctgcaacaa gttccctgca
     2821 atcaaggcca tgttgtccat cgatgtagcc gaagaattcg gaagcggagc tactaacttc
     2881 agcctgctga agcaggctgg cgacgtggag gagaaccctg gacctatggt gagcaagggc
     2941 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3001 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3061 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     3121 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     3181 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3241 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3301 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3361 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3421 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3481 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3541 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3601 cactccaccg gcggcatgga cgagctgtac aagtaactcg agtctagagg gccccttaag
     3661 ggttcgatcc ctaccggtta gtaatgagtt tgatatctcg acaatcaacc tctggattac
     3721 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3781 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3841 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3901 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3961 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     4021 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     4081 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4141 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4201 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4261 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     4321 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4381 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4441 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4501 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4561 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     4621 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4681 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4741 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     4801 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4861 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4921 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4981 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     5041 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5101 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5161 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5221 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5281 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5341 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5401 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5461 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5521 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5581 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5641 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5701 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5761 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5821 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5881 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5941 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     6001 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6061 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6121 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6181 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6241 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6301 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6361 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6421 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6481 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6541 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6601 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6661 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6781 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6841 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6901 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7861 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8641 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8701 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8761 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8821 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8881 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
