LOCUS       V042444                 8228 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042444
VERSION     V042444
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             757..2154
                     /label="VIM(NM_003380)"
                     /note="VIM(NM_003380)"
                     /gene="VIM"
     CDS             2170..2226
                     /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             2227..2937
                     /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    3004..3592
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3658..3706
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3908..4336
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4350..4679
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4746..5540
                     /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    5716..5837
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5886..5902)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5910..5926
                     /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(5934..5964)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5979..6000
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6288..6876)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7047..7907)
                     /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(7908..8012)
                     /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 gccaccatgt ccaccaggtc cgtgtcctcg
      781 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg
      841 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc
      901 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg
      961 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg
     1021 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag
     1081 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat
     1141 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac
     1201 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa
     1261 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa
     1321 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag
     1381 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa
     1441 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag
     1501 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc
     1561 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa
     1621 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc
     1681 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt
     1741 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa
     1801 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag
     1861 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat
     1921 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag
     1981 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg
     2041 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa
     2101 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaagaattc
     2161 ggaagcggag ctactaactt cagcctgctg aagcaggctg gcgacgtgga ggagaaccct
     2221 ggacctatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc
     2281 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
     2341 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc
     2401 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
     2461 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
     2521 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
     2581 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     2641 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     2701 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     2761 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     2821 aacgtcaaca tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag
     2881 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaactc
     2941 gagtctagag ggccccttaa gggttcgatc cctaccggtt agtaatgagt ttgatatctc
     3001 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3061 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3121 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3181 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3241 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3301 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3361 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3421 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3481 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3541 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     3601 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     3661 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     3721 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     3781 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     3841 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     3901 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3961 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4201 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4261 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4321 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4381 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4441 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4501 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4561 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     4621 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4681 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     4741 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4801 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4861 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4921 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4981 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5041 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5101 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5161 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5221 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5281 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5341 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5401 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5461 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5521 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     5581 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5641 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5701 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5761 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5821 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5881 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5941 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6001 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6061 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6481 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6961 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7021 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7081 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7141 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7201 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7261 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7321 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7381 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7441 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7501 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7561 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7621 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7681 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7741 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7801 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7861 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7921 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7981 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8041 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8101 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8161 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8221 tgcttagg
//
