LOCUS       V047479                 8529 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047479
VERSION     V047479
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..2455
                     /label="SLC1A7(NM_006671)"
                     /note="SLC1A7(NM_006671)"
                     /gene="SLC1A7"
     CDS             2471..2527
                     /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             2528..3238
                     /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    3305..3893
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3959..4007
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4209..4637
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4651..4980
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5047..5841
                     /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    6017..6138
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6187..6203)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6211..6227
                     /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(6235..6265)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6280..6301
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6589..7177)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7348..8208)
                     /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(8209..8313)
                     /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 ccaccatggt
      781 gccgcatgcc atcttggcac gggggaggga cgtgtgcagg cggaatggac tcctcatcct
      841 gtctgtgctg tctgtcatcg tgggctgcct cctcggcttc ttcttgagga cccggcgcct
      901 ctcaccacag gaaattagtt acttccagtt ccctggagag ctcctgatga ggatgctgaa
      961 gatgatgatc ctgccactgg tggtctccag cttgatgtcc ggacttgcct ccctggatgc
     1021 caagacctct agccgcctgg gcgtcctcac cgtggcgtac tacctgtgga ccaccttcat
     1081 ggctgtcatc gtgggcatct tcatggtctc catcatccac ccaggcagcg cggcccagaa
     1141 ggagaccacg gagcagagtg ggaagcccat catgagctca gccgatgccc tgttggacct
     1201 catccggaac atgttcccag ccaacctagt agaagccaca ttcaaacagt accgcaccaa
     1261 gaccacccca gttgtcaagt cccccaaggt ggcaccagag gaggcccctc ctcggcggat
     1321 cctcatctac ggggtccagg aggagaatgg ctcccatgtg cagaacttcg ccctggacct
     1381 gaccccgccg cccgaggtcg tttacaagtc agagccgggc accagcgatg gcatgaatgt
     1441 gctgggcatc gtcttcttct ctgccaccat gggcatcatg ctgggccgca tgggtgacag
     1501 cggggccccc ctggtcagct tctgccagtg cctcaatgag tcggtcatga agatcgtggc
     1561 ggtggctgtg tggtatttcc ccttcggcat tgtgttcctc attgcgggta agatcctgga
     1621 gatggacgac cccagggccg tcggcaagaa gctgggcttc tactcagtca ccgtggtgtg
     1681 cgggctggtg ctccacgggc tctttatcct gcccctgctc tacttcttca tcaccaagaa
     1741 gaatcccatc gtcttcatcc gtggcatcct gcaggctctg ctcatcgcgc tggccacctc
     1801 ctccagctca gccacactgc ccatcacctt caagtgcctg ctggagaaca accacatcga
     1861 ccggcgcatc gctcgcttcg tgctgcccgt gggtgccacc atcaacatgg acggcactgc
     1921 gctctacgag gctgtggccg ccatcttcat cgcccaggtc aacaactacg agctggactt
     1981 tggccagatc atcaccatca gtatcacagc cactgcagcc agcattgggg cagctggcat
     2041 cccccaggcc ggcctcgtca ccatggtcat cgtgctcacc tccgtgggac tgcccaccga
     2101 tgacatcacc ctcatcattg ccgttgactg ggctctggac cgtttccgca ccatgattaa
     2161 cgtgctgggt gatgcgctgg cagcggggat catggcccat atatgtcgga aggattttgc
     2221 ccgggacaca ggcaccgaga aactgctgcc ctgcgagacc aagccagtga gcctccagga
     2281 gatcgtggca gcccagcaga atggctgtgt gaagagtgta gccgaggcct ccgagctcac
     2341 cctgggcccc acctgccccc accacgtccc cgttcaagtg gagcaggatg aggagctgcc
     2401 cgctgcgagt ctgaaccact gcaccatcca gatcagtgag ctggagacca atgtcgaatt
     2461 cggaagcgga gctactaact tcagcctgct gaagcaggct ggcgacgtgg aggagaaccc
     2521 tggacctatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2581 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2641 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2701 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2761 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     2821 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     2881 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     2941 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     3001 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     3061 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     3121 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3181 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaact
     3241 cgagtctaga gggcccctta agggttcgat ccctaccggt tagtaatgag tttgatatct
     3301 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3361 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3421 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3481 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3541 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3601 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3661 gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct
     3721 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3781 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3841 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa
     3901 cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     3961 gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt
     4021 tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc
     4081 ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc
     4141 agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg
     4201 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4261 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
     4381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4501 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4561 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
     4621 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4681 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4741 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4801 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4861 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4921 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4981 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5041 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5221 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     5281 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     5341 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5401 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5461 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5521 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5581 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5641 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5701 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5761 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5821 cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac
     5881 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5941 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     6001 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     6061 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     6121 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     6181 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6241 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6301 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6361 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6421 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6481 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6541 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6601 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6661 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6721 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6781 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6841 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6901 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6961 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7021 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7081 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7141 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7201 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7261 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7321 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7381 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7441 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7501 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7561 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7621 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7681 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7741 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7801 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7861 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7921 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7981 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8041 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8101 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8161 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8221 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8281 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8341 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
     8401 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     8461 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     8521 ctgcttagg
//
