LOCUS       V026487                 7048 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026487
VERSION     V026487
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             922..987
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             991..2568
                     /label="Slc3a2(NM_008577)"
                     /note="Slc3a2(NM_008577)"
                     /gene="Slc3a2"
     polyA_signal    2648..2872
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2918..3346
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3360..3689
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3756..4550
                     /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    4724..4845
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4894..4910)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4918..4934
                     /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(4942..4972)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4987..5008
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5296..5881)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6052..6912)
                     /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(6913..7017)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      961 atcgactaca aggatgacga tgacaagctt atgagccagg acaccgaagt ggacatgaaa
     1021 gatgtggagc tgaacgagct agaaccggag aagcagccca tgaatgcagc ggacggggcg
     1081 gcggccgggg agaagaacgg tctggtgaag atcaaggtgg cggaggacga gacggaggcc
     1141 ggggtcaagt tcaccggctt atccaaggag gagctactga aggtagcggg cagccctggc
     1201 tgggtgcgca cccgctgggc gctgctgctg ctcttctggc tcggttggct gggcatgctg
     1261 gcgggcgccg tggttatcat cgttcgggcg ccgcgctgcc gtgagctgcc tgtacagagg
     1321 tggtggcaca agggcgccct ctaccgcatc ggcgaccttc aggcctttgt aggccgggat
     1381 gcgggaggca tagctggtct gaagagccat ctggagtact tgagcaccct gaaggtgaag
     1441 ggcctggtgt taggcccaat tcacaagaac cagaaggatg aaatcaatga aaccgacctg
     1501 aaacagatta atcccacttt gggctcccag gaagatttta aagaccttct acaaagtgcc
     1561 aagaaaaaga gcattcacat cattttggac ctcactccca actaccaggg ccagaatgcg
     1621 tggttcctcc ctgctcaggc tgacattgta gccaccaaaa tgaaggaagc tctgagttct
     1681 tggttgcagg acggtgtgga tggtttccaa ttccgggatg tgggaaagct gatgaatgca
     1741 cccttgtact tggctgagtg gcagaatatc accaagaact taagtgagga caggcttttg
     1801 attgcaggga ctgagtcctc tgacctgcag caaattgtca acatacttga atccaccagc
     1861 gacctgctgt tgaccagctc ctacctgtca aattccactt tcactgggga gcgtactgaa
     1921 tccctagtca ctaggttttt gaatgccact ggcagccaat ggtgcagctg gagtgtgtcg
     1981 caagcaggac tcctcgcaga ctttataccg gaccatcttc tccgactcta ccagctgctg
     2041 ctcttcactc tgccagggac tcctgttttt agctacgggg atgagcttgg ccttcagggt
     2101 gcccttcctg gacagcctgc gaaggcccca ctcatgccgt ggaatgagtc cagcatcttt
     2161 cacatcccaa gacctgtaag cctcaacatg acagtgaagg gccagaatga agaccctggc
     2221 tccctcctta cccagttccg gcggctgagt gaccttcggg gtaaggagcg ctctctgttg
     2281 cacggtgact tccatgcact gtcttcctca cctgacctct tctcctacat acgacactgg
     2341 gaccagaatg agcgttacct ggtggtgctc aacttccgag attcgggccg gtcagccagg
     2401 ctaggggcct ccaacctccc tgctggcata agcctgccag ccagcgctaa acttttgctt
     2461 agtaccgaca gtgcccggca aagccgtgag gaggacacct ccctgaagct ggaaaacctg
     2521 agcctgaatc cttatgaggg cttgctgtta cagttcccct ttgtggcctg agaattctgc
     2581 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     2641 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2701 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2761 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2821 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2881 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2941 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3001 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3061 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3121 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3181 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3241 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3301 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3361 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3421 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3481 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3541 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3601 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3661 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3721 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3781 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3841 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3901 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3961 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4021 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4081 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4141 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4201 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4261 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4321 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4381 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4441 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4501 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4561 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4621 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4681 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4741 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4801 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4861 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4921 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4981 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5041 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5101 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5161 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5221 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5281 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5341 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5401 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5461 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5521 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5581 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5641 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5701 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5761 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5821 cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6001 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6061 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6121 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6181 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6241 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6301 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6361 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6421 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6481 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6541 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6601 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6661 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6721 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6781 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6841 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6901 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6961 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7021 atttccccga aaagtgccac ctgacgtc
//
