LOCUS       V047605                 8700 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047605
VERSION     V047605
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"
     CDS             776..2626
                     /label="SLC6A2(NM_001172501)"
                     /note="SLC6A2(NM_001172501)"
                     /gene="SLC6A2"
     CDS             2642..2698
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2699..3409
                     /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"
     misc_feature    3476..4064
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4130..4178
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4380..4808
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4822..5151
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5218..6012
                     /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    6188..6309
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6358..6374)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6382..6398
                     /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(6406..6436)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6451..6472
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6760..7348)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7519..8379)
                     /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(8380..8484)
                     /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 gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatccag ccaccatgct
      781 tctggcgcgg atgaacccgc aggtgcagcc cgagaacaac ggggcggaca cgggtccaga
      841 gcagcccctt cgggcgcgca aaactgcgga gctgctggtg gtgaaggagc gcaacggcgt
      901 ccagtgcctg ctggcgcccc gcgacggcga cgcgcagccc cgggagacct ggggcaagaa
      961 gatcgacttc ctgctgtccg tagtcggctt cgcagtggac ctggccaacg tgtggcgctt
     1021 cccctacctc tgctacaaga acggcggcgg tgccttcttg atcccgtaca cactgttcct
     1081 tatcatcgcg gggatgcccc tgttctacat ggagctggct ctgggacagt acaaccggga
     1141 gggggctgcc accgtttgga aaatctgccc attcttcaaa ggcgttggct atgctgtcat
     1201 cctgatcgcc ctgtacgttg gcttctacta caacgtcatc atcgcctggt cactctacta
     1261 cctcttctcc tccttcaccc tcaacctgcc ctggaccgac tgtggccaca cctggaacag
     1321 ccccaactgt accgacccca agctcctcaa tggctccgtg cttggcaacc acaccaagta
     1381 ctccaagtac aagttcacgc cggcagccga gttttatgag cgtggtgtcc tgcaccttca
     1441 cgagagcagc gggattcatg acatcggcct gccccagtgg cagctcttgc tctgtctgat
     1501 ggtcgtcgtc atcgtcttgt attttagcct ctggaaaggg gtgaagacat caggaaaggt
     1561 ggtgtggatc acagccacgc tgccttactt cgtgctgttc gtgctcctgg tccatggcgt
     1621 cacgctgccc ggagcctcca atggcatcaa tgcctacctg cacatcgact tctaccgctt
     1681 gaaagaggcc acggtatgga ttgatgccgc aactcagata tttttttcct tgggggctgg
     1741 atttggagta ttgattgcat ttgccagtta caacaaattt gacaacaact gttacaggga
     1801 tgccctgctg accagcagca tcaactgtat caccagcttc gtctctgggt tcgccatctt
     1861 ctccatcctt ggttacatgg cccatgaaca caaggtcaac attgaggatg tggccacaga
     1921 aggagctggc ctagtgttca tcctgtatcc agaggccatt tctaccctgt ctggatctac
     1981 attctgggct gttgtgtttt tcgtcatgct cctggcgctg ggccttgaca gctcaatggg
     2041 aggcatggag gctgtcatca cgggcctggc agatgacttc caggtcctga agcgacaccg
     2101 gaaactcttc acatttggcg tcaccttcag cactttcctt ctcgccctgt tctgcataac
     2161 caagggtgga atttacgtct tgaccctcct ggacaccttt gctgcgggca cctccatcct
     2221 ttttgctgtc ctcatggaag ccatcggagt ttcctggttt tatggagtgg acaggttcag
     2281 caacgacatc cagcagatga tggggttcag gccgggtcta tactggagac tgtgctggaa
     2341 gttcgtcagt cctgccttcc tcctgttcgt ggttgtggtc agcatcatca acttcaagcc
     2401 actcacctac gacgactaca tcttcccgcc ctgggccaac tgggtggggt ggggcatcgc
     2461 cctgtcctcc atggtcctgg tgcccatcta cgtcatctat aagttcctca gcacgcaggg
     2521 ctctctttgg gagagactgg cctatggcat cacgccagag aacgagcacc acctggtggc
     2581 tcagagggac atcagacagt tccagttgca acactggctg gccatcgaat tcggaagcgg
     2641 agctactaac ttcagcctgc tgaagcaggc tggcgacgtg gaggagaacc ctggacctat
     2701 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     2761 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     2821 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     2881 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     2941 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3001 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3061 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3121 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3181 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3241 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3301 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3361 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaac tcgagtctag
     3421 agggcccctt aagggttcga tccctaccgg ttagtaatga gtttgatatc tcgacaatca
     3481 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3541 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3601 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3661 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3721 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3781 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3841 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     3901 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3961 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4021 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     4081 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4141 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4201 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4261 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4321 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     4381 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4441 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4501 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     4561 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4621 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4681 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4741 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4801 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4861 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4921 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4981 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5041 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5101 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5161 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5221 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5281 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5341 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5401 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     5461 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     5521 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     5581 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     5641 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     5701 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     5761 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     5821 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5881 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5941 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6001 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     6061 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6121 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6181 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6241 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6301 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6361 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6421 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6481 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6541 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6601 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6661 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6721 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6781 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6841 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6901 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     6961 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7021 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7081 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7141 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7201 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7261 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7321 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7381 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7441 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7501 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7561 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7621 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7681 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7741 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     7801 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     7861 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     7921 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     7981 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8041 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8101 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8161 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8221 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8281 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8341 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8401 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8461 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     8521 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     8581 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     8641 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
