LOCUS       V041723                 8169 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041723
VERSION     V041723
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             887..2938
                     /label="TWNK(NM_021830)"
                     /note="TWNK(NM_021830)"
                     /gene="TWNK"
     CDS             2975..3685
                     /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    3736..3960
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4006..4434
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4448..4777
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4844..5638
                     /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    5812..5933
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5982..5998)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6006..6022
                     /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(6030..6060)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6075..6096
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6384..6969)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7140..8000)
                     /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(8001..8105)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcaac ttaaggatcc gccaccatgt gggtcctcct
      901 ccgaagtggg taccccctcc gtatcttgtt acccctgcgt ggggagtgga tgggtcggag
      961 gggcctgccc cgaaacttgg ccccaggccc tcctcgcaga cgttacagga aggagactct
     1021 ccaagccttg gatatgccag tgttgcctgt aactgcaact gaaatccgcc agtatttgcg
     1081 ggggcatggg atccccttcc aggatggtca cagttgcctg cgggcactga gcccctttgc
     1141 agagtcttca cagctcaaag gccagactgg tgttaccact tccttcagcc tcttcattga
     1201 caagaccaca ggccactttc tctgcatgac cagcctagca gaagggagct gggaagactt
     1261 ccaggccagc gtggaggggc gaggggatgg ggccagggag gggtttctgc ttagcaaggc
     1321 accagaattt gaggacagcg aggaggtccg gaggatctgg aaccgagcaa tacctctctg
     1381 ggagctgcct gatcaggagg aggttcagct ggctgataca atgtttggcc ttaccaaggt
     1441 tacagatgac acactcaagc gtttcagtgt gcgatatctg cgacctgctc gcagtcttgt
     1501 cttcccttgg ttctcccctg ggggctcagg attacgaggc ctgaagctcc tagaggctaa
     1561 atgccagggg gatggagtga gctacgagga aaccactatt ccccgaccca gcgcctacca
     1621 caatctgttt ggattaccac tgattagtcg tcgagatgct gaggtggtac tgacgagtcg
     1681 tgagcttgac agcctggcct tgaaccagtc cacggggctg cctaccctta ctctaccccg
     1741 aggaacgacc tgcttacccc ctgccttact cccttacctg gaacagttcc ggcggattgt
     1801 attctggttg ggggatgacc ttcggtcctg ggaagccgcc aagttgtttg cacgaaaact
     1861 gaaccccaaa cgatgcttct tggtgcgacc aggagaccag caaccccgtc ccctggaggc
     1921 cctgaacgga ggcttcaatc tttctcgtat tcttcgtacc gccctgcctg cctggcacaa
     1981 gtccatcgta tctttccggc agcttcggga ggaggtgcta ggagaactgt caaatgtgga
     2041 gcaagcagct ggcctccgct ggagccgctt tccagacctc aatcgtatct tgaagggaca
     2101 tcgaaagggc gagctgacgg tcttcacagg gccaacaggc agtggaaaga cgacattcat
     2161 cagtgagtat gccctggatt tgtgttccca gggggtgaac acactgtggg gtagcttcga
     2221 gatcagcaat gtgagactag cccgggtcat gctgacacag tttgccgagg ggcggctgga
     2281 agatcaactg gacaaatatg atcactgggc tgaccgcttt gaggacctgc ccctctattt
     2341 catgactttc catggacagc aaagcatcag gactgtaata gatacaatgc aacatgcagt
     2401 ctacgtctat gacatttgtc atgtgatcat cgacaacctg cagttcatga tgggtcacga
     2461 gcagctgtcc acagacagga tcgcagctca agactacatc atcggggtct ttcggaagtt
     2521 tgcaacagac aataactgcc atgtgacact ggtcattcac ccccggaaag aggatgatga
     2581 caaggaactg cagacagcgt ccatttttgg ctcagccaaa gcaagccagg aagcagacaa
     2641 tgttctgatc ctgcaggaca ggaagctggt aaccgggcca gggaaacggt atctgcaggt
     2701 gtccaagaac cgctttgatg gagatgtagg tgtcttcccg cttgagttca acaagaactc
     2761 cctcaccttc tccattccac caaagaacaa ggcccggctc aagaagatca aggatgacac
     2821 tggaccagtg gccaaaaagc cctcttctgg caaaaagggg gctacgacac agaactctga
     2881 gatttgctca ggccaggccc ccactcccga ccagccagac acctccaagc gttcaaagga
     2941 attcggtgga ggcggatcag gaggcggtgg atctatggtg agcaagggcg aggaggataa
     3001 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     3061 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     3121 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
     3181 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
     3241 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     3301 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     3361 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     3421 ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag
     3481 gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa
     3541 gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca
     3601 caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg
     3661 cggcatggac gagctgtaca agtaggcggc cgctcgagtc tagagggccc gtttaaaccc
     3721 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     3781 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     3841 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3901 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3961 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     4021 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4081 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4141 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4201 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4261 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4321 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4381 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     4441 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4501 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4621 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4681 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4741 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4801 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4861 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4921 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4981 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5041 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5101 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5161 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5221 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5281 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5341 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5401 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5461 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5521 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5581 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5641 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5701 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5761 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5821 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5881 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5941 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6001 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6061 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6121 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6181 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6241 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6301 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6361 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6421 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6481 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6541 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6601 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6661 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6721 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6781 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6841 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6901 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6961 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7021 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7081 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7141 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7201 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7261 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7321 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7381 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7441 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7501 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7561 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7621 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7681 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7741 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7801 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7861 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7921 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7981 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8041 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8101 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8161 ccctatggt
//
