LOCUS       V022506                 6737 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022506
VERSION     V022506
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             908..1615
                     /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             1622..2257
                     /label="BOK(NM_032515)"
                     /note="BOK(NM_032515)"
                     /gene="BOK"
     polyA_signal    2304..2528
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2574..3002
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3016..3345
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3412..4206
                     /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    4380..4501
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4550..4566)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4574..4590
                     /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(4598..4628)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4643..4664
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4952..5537)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5708..6568)
                     /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(6569..6673)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
      961 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     1021 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     1081 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     1141 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     1201 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     1261 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     1321 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     1381 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     1441 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     1501 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     1561 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggaatt
     1621 catggaggtg ctgcggcgct cctcggtctt cgccgccgag atcatggacg cctttgaccg
     1681 ctcgcccaca gacaaggagc tggtggccca ggccaaggcg ctgggccggg agtacgtgca
     1741 cgcgcggctg ctgcgcgccg gcctctcctg gagcgcgccc gagcgtgccg cgccggtccc
     1801 gggacgcctg gctgaggtgt gcgcggtgct gctgcgcctg ggcgatgagc tggagatgat
     1861 ccggcccagc gtctaccgca acgtggcgcg tcagctgcac atctccctgc agtctgagcc
     1921 tgtggtgacc gatgcgttcc tggccgtggc tggccacatc ttctctgcag gcatcacgtg
     1981 gggcaaggtg gtgtccctgt atgcggtggc cgcggggctg gccgtggact gtgtgaggca
     2041 ggcccagcct gccatggtcc acgccctcgt ggactgcctg ggggagttcg tgcgcaagac
     2101 cctggcaacc tggctgcgga gacgcggcgg atggactgat gtcctcaagt gtgtggtcag
     2161 cacagaccct ggcctccgct cccactggct ggtggctgca ctctgcagct tcggccgctt
     2221 cctgaaggct gccttcttcg tgctgctgcc agagagatga ctcgagtcta gagggcccgt
     2281 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     2341 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     2401 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     2461 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     2521 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     2581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2701 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2761 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2881 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2941 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3001 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3061 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3121 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3181 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     3301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     3361 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     3421 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     3481 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     3541 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3601 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3661 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3721 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3781 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3841 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3901 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3961 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4021 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4081 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     4141 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     4201 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     4261 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4321 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4381 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4441 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4501 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4561 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4621 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4681 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4741 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4801 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4861 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4921 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4981 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5041 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5101 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5161 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5221 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5281 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5341 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5401 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5461 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     5521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6721 tcccgatccc ctatggt
//
