LOCUS       V030582                 7001 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030582
VERSION     V030582
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        61..364
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        365..568
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             624..2924
                     /label="Top1(NM_009408)"
                     /note="Top1(NM_009408)"
                     /gene="Top1"
     CDS             2958..3668
                     /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"
     polyA_signal    3789..3910
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      complement(3917..4372)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4399..4503
                     /gene="bla"
                     /label="AmpR promoter"
     promoter        4505..4862
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4897..5691
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5923..5970
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      6299..6887
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcc accatgagcg gggaccatct ccacaacgat tcccagatcg
      661 aagcggattt ccgattgaat gattctcaca aacacaaaga taaacacaaa gatcgagaac
      721 accggcataa ggagcacaag aaggataagg ataaggaccg ggaaaagtct aagcatagca
      781 acagtgaaca taaagattct gaaaagaaac acaaagagaa agagaaaacc aaacacaaag
      841 atggcagctc agaaaaacat aaagacaaac ataaagacag agacaaggaa agacgaaagg
      901 aggaaaagat tagagctgct ggggatgcaa aaataaagaa ggaaaaggaa aatggcttct
      961 ctagtccgcc acgaattaaa gatgaacctg aggatgatgg ctattttgct cctccaaaag
     1021 aggatattaa gccattgaag aggcttcgag atgaagatga tgctgattat aaacctaaga
     1081 aaattaagac agaagatatc aagaaggaga agaaaagaaa atcagaagag gaagaggatg
     1141 gtaaacttaa aaagcccaag aataaagata aagataaaaa agttgctgag ccagataata
     1201 agaaaaagaa gccgaaaaag gaagaggaac agaagtggaa atggtgggaa gaagaacgtt
     1261 atccagaagg catcaaatgg aaattcctag agcataaagg gcctgtcttc gctccaccat
     1321 atgagcctct gccagagagt gtcaagtttt actatgatgg taaagttatg aagctgagtc
     1381 ctaaagcaga agaagtagct acattctttg caaaaatgct tgaccacgaa tatactacta
     1441 aggaaatatt caggaaaaat ttctttaaag attggagaaa ggaaatgact aatgacgaaa
     1501 aaaatacgat taccaaccta agcaaatgcg actttacaca gatgagccag tatttcaaag
     1561 cgcagtcaga agctcggaaa cagatgagca aggaagaaaa attgaaaatc aaagaagaaa
     1621 atgaaaagtt attgaaagaa tacggctttt gtgtaatgga taatcataga gagcggattg
     1681 ccaacttcaa gatagagcct ccggggcttt tccgaggccg agggaaccac cccaagatgg
     1741 gtatgctgaa gagaaggatc atgcctgagg acatcatcat caactgtagc aaagacgcaa
     1801 aggttccttc tccccctcct ggacataaat ggaaggaagt ccgacatgat aacaaggtta
     1861 cttggctggt ctcctggaca gagaatatcc aaggttctat caaatatatc atgctgaatc
     1921 ccagttcacg aatcaagggt gagaaagact ggcagaagta tgagactgcg cggcgtctga
     1981 agaagtgtgt ggacaagatc cgaaatcagt atcgggaaga ctggaagtcc aaagagatga
     2041 aagttcgtca gagagctgta gcactgtact tcattgataa gcttgctctg agagcaggca
     2101 atgagaagga agaaggagag acggcagaca ctgtgggctg ctgttcactt agagtggaac
     2161 acatcaattt acacccagaa ttggatggtc aggagtatgt ggtggaattt gactttcctg
     2221 ggaaggactc aatcagatac tataacaaag tcccagttga gaaacgagtt tttaaaaact
     2281 tacaactgtt tatggagaac aagcagcctg aggacgatct ttttgatcga cttaatactg
     2341 gtattctaaa taaacatctt caggatctca tggagggctt gactgccaag gtgttccgta
     2401 cctacaatgc ctccatcaca ctacagcagc agcttaaaga gctcacagcc cctgatgaga
     2461 atgtaccagc gaagattcta tcttataacc gtgccaatcg agctgttgca attctttgta
     2521 accaccagag ggcgccacca aagacctttg agaagtcaat gatgaacttg cagtctaaga
     2581 ttgatgccaa gaaagatcag ttagcagatg ctcgaaggga cctgaaaagt gctaaggctg
     2641 atgccaaagt catgaaggat gcaaagacca agaaggtagt agagtcaaaa aagaaggctg
     2701 tacagagact agaagagcag ttgatgaagc tggaagttca agccacagat cgagaggaga
     2761 acaaacaaat tgccttgggg acctccaaac tcaattatct ggaccctagg atcacagtgg
     2821 cttggtgcaa aaagtggggg gtcccaattg agaagattta caacaaaacc cagagggaga
     2881 agtttgcttg ggccattgat atgactgatg aggactacga gtttggaggt ggaggttcgg
     2941 atccaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
     3001 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
     3061 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
     3121 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
     3181 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
     3241 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
     3301 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     3361 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     3421 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     3481 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     3541 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     3601 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     3661 acaagtagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     3721 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3781 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3841 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3901 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3961 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     4021 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4081 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4141 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4201 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4261 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4321 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     4381 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4441 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4501 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4561 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4621 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4681 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4741 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4801 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4861 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4921 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4981 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5041 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     5101 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5161 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5221 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5281 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5341 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5401 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     5461 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5521 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5581 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5641 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5701 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5761 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5821 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5881 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5941 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     6001 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     6061 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     6121 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     6181 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     6241 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     6301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     6361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     6421 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     6481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     6541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     6601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     6661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     6721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6901 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6961 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
