LOCUS       V045956                 7986 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045956
VERSION     V045956
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2666
                     /label="Slc6a7(NM_201353)"
                     /note="Slc6a7(NM_201353)"
                     /gene="Slc6a7"
     CDS             2673..2738
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2762..3350
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3416..3464
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3666..4094
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4108..4437
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4504..5298
                     /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    5474..5595
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5644..5660)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5668..5684
                     /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(5692..5722)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5737..5758
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6046..6634)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6805..7665)
                     /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(7666..7770)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgaa gaagctccaa gaagctcacc
      781 tccgcaagcc catcacccca gacctgctga tgactcccag tgaccagggc gatgtggacc
      841 tggatgtaga cttcgcagca gacaggggca actggacggg caagctggac ttcctgctgt
      901 cctgcatcgg ctactgcgtg ggcttgggaa atgtctggcg ctttccctac cgagcctaca
      961 ccaatggagg cggagccttc ctcgtgccct acttcctcat gctggccatc tgtggaatcc
     1021 ccctcttctt tcttgagctc tccctgggcc agttctccag cctgggaccc ctggctgtct
     1081 ggaaaatcag ccctctcttt aaaggtgcag gtgcggccat gctgctcatt gtaggcctgg
     1141 tggccattta ctacaacatg attatcgcct acgtcctctt ctacctcttc gcctccctca
     1201 ccagcaattt gccctgggag cactgtggca actggtggaa cacggagctt tgcctggagc
     1261 acagaggccc caagagtggt aatggggtgc tgcctcttaa cctcagcagc accgtcagcc
     1321 ccagtgagga gtactggagc cgctatgtcc tgcacataca gggcagccag ggcatcggcc
     1381 gaccggggga gattcgctgg aacctctgcc tctgcctgct gctggcctgg gtcatcgtgt
     1441 tcctctgtat cttgaagggg gtgaagtcct cgggcaaggt ggtgtatttc acggccacct
     1501 ttccctacct catcctcctc atgctgctgg ttcgaggagt gacccttccc ggggcctgga
     1561 agggcatcca gttctatctc accccccagt tccaccacct gttatcttca aaggtgtgga
     1621 ttgaagctgc tcttcagatc ttttattctc taggagtggg ctttgggggt ctcctcacct
     1681 ttgcttccta caacacattc catcagaaca tctacagaga caccttcatt gtcactctgg
     1741 gcaatgccat cactagcatc ctggctggct ttgctatctt ctcggtgctg ggctacatgt
     1801 ctcaggagct gggtgtgccc gtggaccaag tggccaaagc aggccctggc ctggcctttg
     1861 ttgtctaccc acaggccatg accatgttgc ctctgtcacc tttctggtcc ttcctcttct
     1921 tcttcatgct tctgacgctt ggcctggata gccagtttgc ctttctagaa accatagtga
     1981 ctgcagtgac tgatgagttc ccatactacc tgcggcccaa gaaggcagtg ttctcaggac
     2041 tcatctgtgt ggccatgtac ctgatgggac tgattctcac cactgatggg ggtatgtact
     2101 ggctggtcct tttggatgac tacagcgcca gcttcggact catggtggtg gtgatcacca
     2161 cgtgcctcgc tgtcacccgg gtatatggca tccagcggtt ctgccgagat atccacatga
     2221 tgctaggctt caagccaggc ctctacttca gggcctgctg gctgtttctg tctccagcta
     2281 cactcttggc cttgctggtg tacagcatcg tcaagtacca gccctcagaa tatggcagct
     2341 atcgcttccc cgcctgggcc gagctgctag gcatcctgat gggtctgctc tcttgcctca
     2401 tgatcccagc tggtatgctg gtagctgtgc ttcgagagga gggctcgctc tgggagagac
     2461 ttcagcaagc cagtcggcct gctatggact ggggcccatc tctggaagag aaccggacgg
     2521 gaatgtatgt ggccacactg gctgggagcc agtcaccaaa gccactgatg gtacacatgc
     2581 gcaaatacgg gggcatcacc agcttcgaga acacggccat tgaggtggac cgtgagatcg
     2641 cagaggagga ggaggagtct atgatgggat ccgactacaa agaccatgac ggtgattata
     2701 aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc gatatctcga
     2761 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2821 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2881 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2941 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3001 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3061 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3121 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     3181 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3241 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3301 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaaacgg
     3361 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3421 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3481 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3541 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     3601 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     3661 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3721 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3781 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3841 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3901 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3961 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4021 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4081 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4141 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4201 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4261 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4321 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4381 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4441 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4501 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4561 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4621 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4681 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4741 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4801 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4861 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4921 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4981 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5041 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5101 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5161 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5221 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5281 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5341 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5401 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5461 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5521 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     5581 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     5641 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     5701 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     5761 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     5821 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5881 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5941 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6001 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6061 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6121 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6181 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6241 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6301 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6361 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6421 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6481 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6541 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     6601 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6661 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6721 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6781 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6841 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6901 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6961 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7021 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7081 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7141 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7201 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7261 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7321 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7381 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7441 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7501 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7561 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7621 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7681 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7741 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7801 cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat ctgctctgat
     7861 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     7921 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     7981 cttagg
//
