LOCUS       V047503                 8655 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047503
VERSION     V047503
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..2581
                     /label="SLC6A13(NM_016615)"
                     /note="SLC6A13(NM_016615)"
                     /gene="SLC6A13"
     CDS             2597..2653
                     /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             2654..3364
                     /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    3431..4019
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4085..4133
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4335..4763
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4777..5106
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5173..5967
                     /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    6143..6264
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6313..6329)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6337..6353
                     /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(6361..6391)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6406..6427
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6715..7303)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7474..8334)
                     /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(8335..8439)
                     /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 tagcagggtc tcaggcacaa ccagtaatgg agagacaaaa ccagtgtatc cagtcatgga
      841 aaagaaggag gaagatggca ccctggagcg ggggcactgg aacaacaaga tggagtttgt
      901 gctgtcagtg gctggggaga tcattggctt aggcaacgtc tggaggtttc cctatctctg
      961 ctacaaaaat gggggaggtg ccttcttcat cccctacctc gtcttcctct ttacctgtgg
     1021 cattcctgtc ttccttctgg agacagcact aggccagtac actagccagg gaggcgtcac
     1081 agcctggagg aagatctgcc ccatctttga gggcattggc tatgcctccc agatgatcgt
     1141 catcctcctc aacgtctact acatcattgt gttggcctgg gccctgttct acctcttcag
     1201 cagcttcacc atcgacctgc cctggggcgg ctgctaccat gagtggaaca cagaacactg
     1261 tatggagttc cagaagacca acggctccct gaatggtacc tctgagaatg ccacctctcc
     1321 tgtcatcgag ttctgggagc ggcgggtctt gaagatctct gatgggatcc agcacctggg
     1381 ggccctgcgc tgggagctgg ctctgtgcct cctgctggcc tgggtcatct gctacttctg
     1441 catctggaag ggggtgaagt ccacaggcaa ggtggtgtac ttcacggcca catttcctta
     1501 cctcatgctg gtggtcctgt taattcgagg ggtgacgttg cctggggcag cccaaggaat
     1561 tcagttttac ctgtacccaa acctcacgcg tctgtgggat ccccaggtgt ggatggatgc
     1621 aggcacccag atattcttct ccttcgccat ctgtcttggg tgcctgacag ccctgggcag
     1681 ctacaacaag taccacaaca actgctacag ggactgcatc gccctctgct tcctcaacag
     1741 cggcaccagc tttgtggccg gctttgccat cttctccatc ctgggcttca tgtctcagga
     1801 gcagggggtg cccatttctg aggtggccga gtcaggccct ggcctggctt tcatcgctta
     1861 cccgcgggct gtggtgatgc tgcccttctc tcctctctgg gcctgctgtt tcttcttcat
     1921 ggtcgttctc ctgggactgg atagccagtt tgtgtgtgta gaaagcctgg tgacagcgct
     1981 ggtggacatg taccctcacg tgttccgcaa gaagaaccgg agggaagtcc tcatccttgg
     2041 agtatctgtc gtctccttcc ttgtggggct gatcatgctc acagagggcg gaatgtacgt
     2101 gttccagctc tttgactact atgcggccag tggcatgtgc ctcctgttcg tggccatctt
     2161 cgagtccctc tgtgtggctt gggtttacgg agccaagcgc ttctacgaca acatcgaaga
     2221 catgattggg tacaggccat ggcctcttat caaatactgt tggctcttcc tcacaccagc
     2281 tgtgtgcaca gccacctttc tcttctccct gataaagtac actccgctga cctacaacaa
     2341 gaagtacacg tacccgtggt ggggcgatgc cctgggctgg ctcctggctc tgtcctccat
     2401 ggtctgcatt cctgcctgga gcctctacag actcggaacc ctcaagggcc ccttcagaga
     2461 gagaatccgt cagctcatgt gcccagccga ggacctgccc cagcggaacc cagcaggacc
     2521 ctcggctccc gccaccccca ggacctcact gctcagactc acagagctag agtctcactg
     2581 cgaattcgga agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggagga
     2641 gaaccctgga cctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2701 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2761 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2821 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2881 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2941 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     3001 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     3061 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     3121 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     3181 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     3241 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     3301 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     3361 gtaactcgag tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg
     3421 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     3481 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     3541 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     3601 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     3661 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3721 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3781 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3841 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3901 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3961 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     4021 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     4081 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     4141 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4201 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4261 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     4321 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4381 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     4501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4681 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     4741 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4801 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4861 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4921 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4981 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5041 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5101 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5161 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5221 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5281 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5341 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5401 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5461 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5521 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5581 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5641 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5701 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5761 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5821 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5881 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5941 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     6001 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     6061 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     6121 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     6181 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     6241 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     6301 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     6361 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     6421 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6481 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6541 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6601 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6661 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6721 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6781 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6841 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6901 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6961 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7021 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7081 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7141 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7201 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7261 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7321 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7381 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     7441 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     7501 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     7561 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     7621 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     7681 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7741 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7801 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7861 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7921 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7981 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     8041 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     8101 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     8161 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     8221 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     8281 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     8341 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     8401 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     8461 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     8521 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     8581 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     8641 aagaatctgc ttagg
//
