LOCUS       V025859                 9402 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025859
VERSION     V025859
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             744..1451
                     /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"
     CDS             1500..4082
                     /label="ORC1(NM_004153)"
                     /note="ORC1(NM_004153)"
                     /gene="ORC1"
     CDS             4089..4154
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4178..4766
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4832..4880
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5082..5510
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5524..5853
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5920..6714
                     /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    6890..7011
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7060..7076)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7084..7100
                     /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(7108..7138)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7153..7174
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7462..8050)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8221..9081)
                     /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(9082..9186)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgcc accatggtga gcaagggcga ggaggataac atggccatca
      781 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      841 agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg
      901 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
      961 gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc
     1021 ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga
     1081 cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca
     1141 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg
     1201 agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga
     1261 aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc
     1321 agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact
     1381 acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg
     1441 agctgtacaa gtggatcgga ggagggggaa gcggtggagg cggtagtgag ctcggatcca
     1501 tggcacacta ccccacaagg ctgaagacca gaaaaactta ttcatgggtt ggcaggccct
     1561 tgttggatcg aaaactgcac taccaaacct atagagaaat gtgtgtgaaa acagaaggtt
     1621 gttccaccga gattcacatc cagattggac agtttgtgtt gattgaaggg gatgatgatg
     1681 aaaacccgta tgttgctaaa ttgcttgagt tgttcgaaga tgactctgat cctcctccta
     1741 agaaacgtgc tcgagtacag tggtttgtcc gattctgtga agtccctgcc tgtaaacggc
     1801 atttgttggg ccggaagcct ggtgcacagg aaatattctg gtatgattac ccggcctgtg
     1861 acagcaacat taatgcggag accatcattg gccttgttcg ggtgatacct ttagccccaa
     1921 aggatgtggt accgacgaat ctgaaaaatg agaagacact ctttgtgaaa ctatcctgga
     1981 atgagaagaa attcaggcca ctttcctcag aactatttgc ggagttgaat aaaccacaag
     2041 agagtgcagc caagtgccag aaacccgtga gagccaagag taagagtgca gagagccctt
     2101 cttggacccc agcagaacat gtggccaaaa ggattgaatc aaggcactcc gcctccaaat
     2161 ctcgccaaac tcctacccat cctcttaccc caagagccag aaagaggctg gagcttggca
     2221 acttaggtaa ccctcagatg tcccagcaga cttcatgtgc ctccttggat tctccaggaa
     2281 gaataaaacg gaaagtggcc ttctcggaga tcacctcacc ttctaagaga tctcagcctg
     2341 ataaacttca aaccttgtct ccagctctga aagccccaga gaaaaccaga gagactggac
     2401 tctcttatac tgaggatgac aagaaggctt cacctgaaca tcgcataatc ctgagaaccc
     2461 gaattgcagc ttcgaaaacc atagacatta gagaggagag aacacttacc cctatcagtg
     2521 ggggacagag atcttcagtg gtgccatccg tgattctgaa accagaaaac atcaaaaaga
     2581 gggatgcaaa agaagcaaaa gcccagaatg aagcgacctc tactccccat cgtatccgca
     2641 gaaagagttc tgtcttgact atgaatcgga ttaggcagca gcttcggttt ctaggtaata
     2701 gtaaaagtga ccaagaagag aaagagattc tgccagcagc agagatttca gactctagca
     2761 gtgacgaaga agaggcttcc acaccgcccc ttccaaggag agcacccaga actgtgtcca
     2821 ggaacctgcg atcttccttg aagtcatcct tacataccct cacgaaggtg ccaaagaaga
     2881 gtctcaagcc tagaacgcca cgttgtgccg ctcctcagat ccgtagtcga agcctggctg
     2941 cccaggagcc agccagtgtg ctggaggaag cccgactgag gctgcatgtt tctgctgtac
     3001 ctgagtctct tccctgtcgg gaacaggaat tccaagacat ctacaatttt gtggaaagca
     3061 aactccttga ccataccgga gggtgcatgt acatctccgg tgtccctggg acagggaaga
     3121 ctgccactgt tcatgaagtg atacgctgcc tgcagcaggc agcccaagcc aatgatgttc
     3181 ctccctttca atacattgag gtcaatggca tgaagctgac ggagccccac caagtctatg
     3241 tgcaaatctt gcagaagcta acaggccaaa aagcaacagc caaccatgcg gcagaactgc
     3301 tggcaaagca attctgcacc cgagggtcac ctcaggaaac caccgtcctg cttgtggatg
     3361 agctcgacct tctgtggact cacaaacaag acataatgta caatctcttt gactggccca
     3421 ctcataagga ggcccggctt gtggtcctgg caattgccaa cacaatggac ctgccagagc
     3481 gaatcatgat gaaccgggtg tccagccgac tgggtcttac caggatgtgc ttccagccct
     3541 atacatatag ccagctgcag cagatcctaa ggtcccggct caagcatcta aaggcctttg
     3601 aagatgatgc catccagctg gtagccagga aggtagcagc actgtctgga gatgcacgac
     3661 ggtgcctgga catctgcagg cgtgccacag agatctgtga gttctcccag cagaagcctg
     3721 actcccctgg cctggtcacc atagcccact caatggaagc tgtggatgag atgttttcat
     3781 catcatacat cacggccatc aaaaattcct ctgttctgga acagagcttc ctgagagcca
     3841 tcctcgcaga gttccgtcga tcaggactgg aggaagccac gtttcaacag atatatagtc
     3901 aacatgtggc actgtgcaga atggagggac tgccgtaccc caccatgtca gagaccatgg
     3961 ccgtgtgttc tcacctgggc tcctgtcgcc tcctgcttgt ggagcccagc aggaacgatc
     4021 tgctccttcg ggtgcggctc aacgtcagcc aggatgatgt gctgtatgcg ctgaaagacg
     4081 agctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     4141 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     4201 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4261 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4321 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4381 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4441 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4501 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4561 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4621 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     4681 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     4741 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     4801 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4861 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     4921 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     4981 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5041 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5101 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5161 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5221 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     5281 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5341 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5401 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5461 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5521 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5581 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5641 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5701 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5761 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5821 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5881 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     5941 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6001 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6061 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6121 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6181 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6241 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6301 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6361 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6421 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6481 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6541 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6601 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6661 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6721 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6781 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6841 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6901 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     6961 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7021 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7081 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7621 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     7681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7861 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8221 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8281 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8341 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8401 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8461 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8521 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8581 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8641 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8701 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8761 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8821 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8881 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8941 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9001 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9061 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9121 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9181 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     9241 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     9301 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     9361 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
