LOCUS       V047510                 9240 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047510
VERSION     V047510
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..3166
                     /label="SLC6A5(NM_004211)"
                     /note="SLC6A5(NM_004211)"
                     /gene="SLC6A5"
     CDS             3182..3238
                     /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             3239..3949
                     /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    4016..4604
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4670..4718
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4920..5348
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5362..5691
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5758..6552
                     /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    6728..6849
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6898..6914)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6922..6938
                     /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(6946..6976)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6991..7012
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7300..7888)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8059..8919)
                     /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(8920..9024)
                     /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 ccaccatgga
      781 ttgcagtgct cccaaggaaa tgaataaact gccagccaac agcccggagg cggcggcggc
      841 gcagggccac ccggatggcc catgcgctcc caggacgagc ccggagcagg agcttcccgc
      901 ggctgccgcc ccgccgccgc cacgtgtgcc caggtccgct tccaccggcg cccaaacttt
      961 ccagtcagcg gacgcgcgag cctgcgaggc tgagcggcca ggagtggggt cttgcaaact
     1021 cagtagcccg cgggcgcagg cggcctctgc agctctgcgg gacttgagag aggcgcaagg
     1081 cgcgcaggcc tcgccccctc ccgggagctc cgggcccggc aacgcgctgc actgtaagat
     1141 cccttttctg cgaggcccgg agggggatgc gaacgtgagt gtgggcaagg gcaccctgga
     1201 gcggaacaat acccctgttg tgggctgggt gaacatgagc cagagcaccg tggtgctggc
     1261 cacggatgga atcacgtccg tgctcccggg cagcgtggcc accgttgcca cccaggagga
     1321 cgagcaaggg gatgagaata aggcccgagg gaactggtcc agcaaactgg acttcatcct
     1381 gtccatggtg gggtacgcag tggggctggg caatgtctgg aggtttccct acctggcctt
     1441 ccagaacggg ggaggtgctt tcctcatccc ttacctgatg atgctggctc tggctggatt
     1501 acccatcttc ttcttggagg tgtcgctggg ccagtttgcc agccagggac cagtgtctgt
     1561 gtggaaggcc atcccagctc tacaaggctg tggcatcgcg atgctgatca tctctgtcct
     1621 aatagccata tactacaatg tgattatttg ctatacactt ttctacctgt ttgcctcctt
     1681 tgtgtctgta ctaccctggg gctcctgcaa caacccttgg aatacgccag aatgcaaaga
     1741 taaaaccaaa cttttattag attcctgtgt tatcagtgac catcccaaaa tacagatcaa
     1801 gaactcgact ttctgcatga ccgcttatcc caacgtgaca atggttaatt tcaccagcca
     1861 ggccaataag acatttgtca gtggaagtga agagtacttc aagtactttg tgctgaagat
     1921 ttctgcaggg attgaatatc ctggcgagat caggtggcca ctagctctct gcctcttcct
     1981 ggcttgggtc attgtgtatg catcattggc taaaggaatc aagacttcag gaaaagtggt
     2041 gtacttcacg gccacgttcc cgtatgtcgt actcgtgatc ctcctcatcc gaggagtcac
     2101 cctgcctgga gctggagctg ggatctggta cttcatcaca cccaagtggg agaaactcac
     2161 ggatgccacg gtgtggaaag atgctgccac tcagattttc ttctctttat ctgctgcatg
     2221 gggaggcctg atcactctct cttcttacaa caaattccac aacaactgct acagggacac
     2281 tctaattgtc acctgcacca acagtgccac aagcatcttt gccggcttcg tcatcttctc
     2341 cgttatcggc ttcatggcca atgaacgcaa agtcaacatt gagaatgtgg cagaccaagg
     2401 gccaggcatt gcatttgtgg tttacccgga agccttaacc aggctgcctc tctctccgtt
     2461 ctgggccatc atctttttcc tgatgctcct cactcttgga cttgacacta tgtttgccac
     2521 catcgagacc atagtgacct ccatctcaga cgagtttccc aagtacctac gcacacacaa
     2581 gccagtgttt actctgggct gctgcatttg tttcttcatc atgggttttc caatgatcac
     2641 tcagggtgga atttacatgt ttcagcttgt ggacacctat gctgcctcct atgcccttgt
     2701 catcattgcc atttttgagc tcgtggggat ctcttatgtg tatggcttgc aaagattctg
     2761 tgaagatata gagatgatga ttggattcca gcctaacatc ttctggaaag tctgctgggc
     2821 atttgtaacc ccaaccattt taacctttat cctttgcttc agcttttacc agtgggagcc
     2881 catgacctat ggctcttacc gctatcctaa ctggtccatg gtgctcggat ggctaatgct
     2941 cgcctgttcc gtcatctgga tcccaattat gtttgtgata aaaatgcatc tggcccctgg
     3001 aagatttatt gagaggctga agttggtgtg ctcgccacag ccggactggg gcccattctt
     3061 agctcaacac cgcggggagc gttacaagaa catgatcgac cccttgggaa cctcttcctt
     3121 gggactcaaa ctgccagtga aggatttgga actgggcact cagtgcgaat tcggaagcgg
     3181 agctactaac ttcagcctgc tgaagcaggc tggcgacgtg gaggagaacc ctggacctat
     3241 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3301 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3361 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3421 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3481 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3541 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3601 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3661 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3721 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3781 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3841 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3901 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaac tcgagtctag
     3961 agggcccctt aagggttcga tccctaccgg ttagtaatga gtttgatatc tcgacaatca
     4021 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4081 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4141 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4201 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4261 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4321 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4381 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     4441 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4501 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4561 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa acgggggagg
     4621 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     4681 agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4741 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4801 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4861 cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca
     4921 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4981 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5041 gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag
     5101 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5161 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5221 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5281 agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5341 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5401 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5461 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5521 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5581 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5641 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5701 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5761 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5821 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     5881 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     5941 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6001 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6061 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6121 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6181 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6241 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6301 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6361 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6421 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6481 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6541 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     6601 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6661 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6721 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6781 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6841 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     6901 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6961 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7021 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     7501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7741 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     9061 atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc tgatgccgca
     9121 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     9181 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
//
