LOCUS       V027500                 9374 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027500
VERSION     V027500
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..3300
                     /label="UNC5A(NM_133369)"
                     /note="UNC5A(NM_133369)"
                     /gene="UNC5A"
     CDS             3316..3372
                     /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             3373..4083
                     /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    4150..4738
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4804..4852
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5054..5482
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5496..5825
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5892..6686
                     /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    6862..6983
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7032..7048)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7056..7072
                     /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(7080..7110)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7125..7146
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7434..8022)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8193..9053)
                     /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(9054..9158)
                     /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 caccatggcc
      781 gtccggcccg gcctgtggcc agcgctcctg ggcatagtcc tcgccgcttg gctccgcggc
      841 tcgggtgccc agcagagtgc caccgtggcc aacccagtgc ctggtgccaa cccggacctg
      901 cttccccact tcctggtgga gcccgaggat gtgtacatcg tcaagaacaa gccagtgctg
      961 cttgtgtgca aggccgtgcc cgccacgcag atcttcttca agtgcaacgg ggagtgggtg
     1021 cgccaggtgg accacgtgat cgagcgcagc acagacggga gcagtgggct gcccaccatg
     1081 gaggtccgca ttaatgtctc aaggcagcag gtcgagaagg tgttcgggct ggaggaatac
     1141 tggtgccagt gcgtggcatg gagctcctcg ggcaccacca agagtcagaa ggcctacatc
     1201 cgcatagcct atttgcgcaa gaacttcgag caggagccgc tggccaagga ggtgtccctg
     1261 gagcagggca tcgtgctgcc ctgccgtcca ccggagggca tccctccagc cgaggtggag
     1321 tggctccgga acgaggacct ggtggacccg tccctggacc ccaatgtata catcacgcgg
     1381 gagcacagcc tggtggtgcg acaggcccgc cttgctgaca cggccaacta cacctgcgtg
     1441 gccaagaaca tcgtggcacg tcgccgcagc gcctccgctg ctgtcatcgt ctacgtggac
     1501 ggcagctgga gcccgtggag caagtggtcg gcctgtgggc tggactgcac ccactggcgg
     1561 agccgtgagt gctctgaccc agcaccccgc aacggagggg aggagtgcca gggcactgac
     1621 ctggacaccc gcaactgtac cagtgacctc tgtgtacaca ctgcttctgg ccctgaggac
     1681 gtggccctct atgtgggcct catcgccgtg gccgtctgcc tggtcctgct gctgcttgtc
     1741 ctcatcctcg tttattgccg gaagaaggag gggctggact cagatgtggc tgactcgtcc
     1801 attctcacct caggcttcca gcccgtcagc atcaagccca gcaaagcaga caacccccat
     1861 ctgctcacca tccagccgga cctcagcacc accaccacca cctaccaggg cagtctctgt
     1921 ccccggcagg atgggcccag ccccaagttc cagctcacca atgggcacct gctcagcccc
     1981 ctgggtggcg gccgccacac actgcaccac agctctccca cctctgaggc cgaggagttc
     2041 gtctcccgcc tctccaccca gaactacttc cgctccctgc cccgaggcac cagcaacatg
     2101 acctatggga ccttcaactt cctcgggggc cggctgatga tccctaatac aggaatcagc
     2161 ctcctcatcc ccccagatgc cataccccga gggaagatct atgagatcta cctcacgctg
     2221 cacaagccgg aagacgtgag gttgccccta gctggctgtc agaccctgct gagtcccatc
     2281 gttagctgtg gaccccctgg cgtcctgctc acccggccag tcatcctggc tatggaccac
     2341 tgtggggagc ccagccctga cagctggagc ctgcgcctca aaaagcagtc gtgcgagggc
     2401 agctgggagg atgtgctgca cctgggcgag gaggcgccct cccacctcta ctactgccag
     2461 ctggaggcca gtgcctgcta cgtcttcacc gagcagctgg gccgctttgc cctggtggga
     2521 gaggccctca gcgtggctgc cgccaagcgc ctcaagctgc ttctgtttgc gccggtggcc
     2581 tgcacctccc tcgagtacaa catccgggtc tactgcctgc atgacaccca cgatgcactc
     2641 aaggaggtgg tgcagctgga gaagcagctg gggggacagc tgatccagga gccacgggtc
     2701 ctgcacttca aggacagtta ccacaacctg cgcctatcca tccacgatgt gcccagctcc
     2761 ctgtggaaga gtaagctcct tgtcagctac caggagatcc ccttttatca catctggaat
     2821 ggcacgcagc ggtacttgca ctgcaccttc accctggagc gtgtcagccc cagcactagt
     2881 gacctggcct gcaagctgtg ggtgtggcag gtggagggcg acgggcagag cttcagcatc
     2941 aacttcaaca tcaccaagga cacaaggttt gctgagctgc tggctctgga gagtgaagcg
     3001 ggggtcccag ccctggtggg ccccagtgcc ttcaagatcc ccttcctcat tcggcagaag
     3061 ataatttcca gcctggaccc accctgtagg cggggtgccg actggcggac tctggcccag
     3121 aaactccacc tggacagcca tctcagcttc tttgcctcca agcccagccc cacagccatg
     3181 atcctcaacc tgtgggaggc gcggcacttc cccaacggca acctcagcca gctggctgca
     3241 gcagtggctg gactgggcca gccagacgct ggcctcttca cagtgtcgga ggctgagtgc
     3301 gaattcggaa gcggagctac taacttcagc ctgctgaagc aggctggcga cgtggaggag
     3361 aaccctggac ctatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     3421 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3481 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3541 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3601 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3661 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3721 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3781 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3841 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3901 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3961 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     4021 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     4081 taactcgagt ctagagggcc ccttaagggt tcgatcccta ccggttagta atgagtttga
     4141 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4201 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4261 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4321 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     4381 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     4441 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     4501 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     4561 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     4621 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     4681 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     4741 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4801 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     4861 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     4921 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     4981 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     5041 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5101 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5161 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     5221 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     5281 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5341 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     5401 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     5461 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5521 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5581 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5641 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5701 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5761 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5821 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     5881 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5941 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6001 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6061 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6121 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6181 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6241 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6301 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6361 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     6421 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     6481 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     6541 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     6601 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     6661 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     6721 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6781 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6841 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     6901 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     6961 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     7021 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     7081 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7141 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7201 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7261 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7321 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7381 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7441 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7501 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7561 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7621 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7681 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7741 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7801 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7861 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7921 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7981 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     8041 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     8101 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8161 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8221 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8281 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8341 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8401 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8461 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8521 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8581 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8641 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8701 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8761 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8821 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8881 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8941 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     9001 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     9061 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     9121 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     9181 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     9241 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     9301 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     9361 agaatctgct tagg
//
