LOCUS       V040963                 7808 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040963
VERSION     V040963
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             872..1579
                     /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             1616..3346
                     /label="IGF2BP1(NM_006546)"
                     /note="IGF2BP1(NM_006546)"
                     /gene="IGF2BP1"
     CDS             3353..3379
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3383..3409
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3413..3439
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3446..3472
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3476..3502
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3506..3532
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3570..3794
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3840..4268
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4282..4611
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4678..5277
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5451..5572
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5621..5637)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5645..5661
                     /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(5669..5699)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5714..5735
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6023..6608)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6779..7639)
                     /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(7640..7744)
                     /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 ggctagccac catggtgagc aagggcgagg aggataacat
      901 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      961 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1021 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1081 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1141 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1201 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1261 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1321 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1381 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1441 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1501 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1561 catggacgag ctgtacaagg gaggtggagg tagtggagga ggcggatcaa gcttcatgaa
     1621 caagctttac atcggcaacc tcaacgagag cgtgaccccc gcggacttgg agaaagtgtt
     1681 tgcggagcac aagatctcct acagcggcca gttcttggtc aaatccggct acgccttcgt
     1741 ggactgcccg gacgagcact gggcgatgaa ggccatcgaa actttctccg ggaaagtaga
     1801 attacaagga aaacgcttag agattgaaca ttcggtgccc aaaaaacaaa ggagccggaa
     1861 aattcaaatc cgaaatattc caccccagct ccgatgggaa gtactggaca gcctgctggc
     1921 tcagtatggt acagtagaga actgtgagca agtgaacacc gagagtgaga cggcagtggt
     1981 gaatgtcacc tattccaacc gggagcagac caggcaagcc atcatgaagc tgaatggcca
     2041 ccagttggag aaccatgccc tgaaggtctc ctacatcccc gatgagcaga tagcacaggg
     2101 acctgagaat gggcgccgag ggggctttgg ctctcggggt cagccccgcc agggctcacc
     2161 tgtggcagcg ggggccccag ccaagcagca gcaagtggac atcccccttc ggctcctggt
     2221 gcccacccag tatgtgggtg ccattattgg caaggagggg gccaccatcc gcaacatcac
     2281 aaaacagacc cagtccaaga tagacgtgca taggaaggag aacgcaggtg cagctgaaaa
     2341 agccatcagt gtgcactcca cccctgaggg ctgctcctcc gcttgtaaga tgatcttgga
     2401 gattatgcat aaagaggcta aggacaccaa aacggctgac gaggttcccc tgaagatcct
     2461 ggcccataat aactttgtag ggcgtctcat tggcaaggaa ggacggaacc tgaagaaggt
     2521 agagcaagat accgagacaa aaatcaccat ctcctcgttg caagacctta ccctttacaa
     2581 ccctgagagg accatcactg tgaagggggc catcgagaat tgttgcaggg ccgagcagga
     2641 aataatgaag aaagttcggg aggcctatga gaatgatgtg gctgccatga gcctgcagtc
     2701 tcacctgatc cctggcctga acctggctgc tgtaggtctt ttcccagctt catccagcgc
     2761 agtcccgccg cctcccagca gcgttactgg ggctgctccc tatagctcct ttatgcaggc
     2821 tcccgagcag gagatggtgc aggtgtttat ccccgcccag gcagtgggcg ccatcatcgg
     2881 caagaagggg cagcacatca aacagctctc ccggtttgcc agcgcctcca tcaagattgc
     2941 accacccgaa acacctgact ccaaagttcg tatggttatc atcactggac cgccagaggc
     3001 ccaattcaag gctcagggaa gaatctatgg caaactcaag gaggagaact tctttggtcc
     3061 caaggaggaa gtgaagctgg agacccacat acgtgtgcca gcatcagcag ctggccgggt
     3121 cattggcaaa ggtggaaaaa cggtgaacga gttgcagaat ttgacggcag ctgaggtggt
     3181 agtaccaaga gaccagaccc ctgatgagaa cgaccaggtc atcgtgaaaa tcatcggaca
     3241 tttctatgcc agtcagatgg ctcaacggaa gatccgagac atcctggccc aggttaagca
     3301 gcagcatcag aagggacaga gtaaccaggc ccaggcacgg aggaagctcg agtatccgta
     3361 tgacgttccg gattacgcga cctatccgta cgacgttccg gattacgctt cctacccata
     3421 cgacgtaccg gattacgcgc tcgagtaccc atacgacgta cctgattatg caacttatcc
     3481 ttatgatgta cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg
     3541 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3601 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3661 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3721 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3781 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3841 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3961 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4201 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4261 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4321 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4381 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4441 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4501 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4561 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4621 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4681 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     4741 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     4801 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4861 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4921 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4981 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5041 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5101 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5161 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5221 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagcg
     5281 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5341 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5401 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5461 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5521 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5581 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5641 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5701 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5761 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5821 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5881 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5941 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6001 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6061 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6121 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6181 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6241 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6301 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6361 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6421 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6481 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6541 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7741 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7801 cctatggt
//
