LOCUS       V034208                 7797 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034208
VERSION     V034208
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..2477
                     /label="SLC1A2(NM_004171)"
                     /note="SLC1A2(NM_004171)"
                     /gene="SLC1A2"
     CDS             2484..2549
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2573..3161
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3227..3275
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3477..3905
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3919..4248
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4315..5109
                     /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    5285..5406
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5455..5471)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5479..5495
                     /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(5503..5533)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5548..5569
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5857..6445)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6616..7476)
                     /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(7477..7581)
                     /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 ccaccatggc atctacggaa ggtgccaaca
      781 atatgcccaa gcaggtggaa gtgcgaatgc acgacagtca tcttggctca gaggaaccca
      841 agcaccggca cctgggcctg cgcctgtgtg acaagctggg gaagaatctg ctgctcaccc
      901 tgacggtgtt tggtgtcatc ctgggagcag tgtgtggagg gcttcttcgc ttggcatctc
      961 ccatccaccc tgatgtggtt atgttaatag ccttcccagg ggatatactc atgaggatgc
     1021 taaaaatgct cattctccct ctaatcatct ccagcttaat cacagggttg tcaggcctgg
     1081 atgctaaggc tagtggccgc ttgggcacga gagccatggt gtattacatg tccacgacca
     1141 tcattgctgc agtactgggg gtcattctgg tcttggctat ccatccaggc aatcccaagc
     1201 tcaagaagca gctggggcct gggaagaaga atgatgaagt gtccagcctg gatgccttcc
     1261 tggaccttat tcgaaatctc ttccctgaaa accttgtcca agcctgcttt caacagattc
     1321 aaacagtgac gaagaaagtc ctggttgcac caccgccgga cgaggaggcc aacgcaacca
     1381 gcgctgttgt ctctctgttg aacgagactg tgactgaggt gccggaggag actaagatgg
     1441 ttatcaagaa gggcctggag ttcaaggatg ggatgaacgt cttaggtctg atagggtttt
     1501 tcattgcttt tggcatcgct atggggaaga tgggagatca ggccaagctg atggtggatt
     1561 tcttcaacat tttgaatgag attgtaatga agttagtgat catgatcatg tggtactctc
     1621 ccctgggtat cgcctgcctg atctgtggaa agatcattgc aatcaaggac ttagaagtgg
     1681 ttgctaggca actggggatg tacatggtaa cagtgatcat aggcctcatc atccacgggg
     1741 gcatctttct ccccttgatt tactttgtag tgaccaggaa aaaccccttc tccttttttg
     1801 ctggcatttt ccaagcttgg atcactgccc tgggcaccgc ttccagtgct ggaactttgc
     1861 ctgtcacctt tcgttgcctg gaagaaaatc tggggattga taagcgtgtg actagattcg
     1921 tccttcctgt tggagcaacc attaacatgg atggtacagc cctttatgaa gcggtagccg
     1981 ccatctttat agcccaaatg aatggtgttg tcctggatgg aggacagatt gtgactgtaa
     2041 gcctcacagc caccctggca agcgtcggcg cggccagtat ccccagtgcc gggctggtca
     2101 ccatgctcct cattctgaca gccgtgggcc tgccaacaga ggacatcagc ctgctggtgg
     2161 ctgtggactg gctgctggac aggatgagaa cttcagtcaa tgttgtgggt gactcttttg
     2221 gggctgggat agtctatcac ctctccaagt ctgagctgga taccattgac tcccagcatc
     2281 gagtgcatga agatattgaa atgaccaaga ctcaatccat ttatgatgac atgaagaacc
     2341 acagggaaag caactctaat caatgtgtct atgctgcaca caactctgtc atagtagatg
     2401 aatgcaaggt aactctggca gccaatggaa agtcagccga ctgcagtgtt gaggaagaac
     2461 cttggaaacg tgagaaagga tccgactaca aagaccatga cggtgattat aaagatcatg
     2521 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     2581 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2641 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2701 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2761 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2821 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2881 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2941 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3001 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3061 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3121 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     3181 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     3241 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     3301 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     3361 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     3421 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     3481 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3541 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3601 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     3661 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3721 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3781 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3841 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3901 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3961 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4081 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4141 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4201 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4261 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4321 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4381 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4441 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4501 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4561 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4621 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4681 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4741 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4801 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4861 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4921 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4981 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5041 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5101 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     5161 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5221 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5281 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5341 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5401 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     5461 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5521 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5581 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5641 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5701 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6421 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6541 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6601 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6661 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6721 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6781 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6841 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6901 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6961 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7021 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7081 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7141 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     7201 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     7261 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     7321 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     7381 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     7441 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     7501 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7561 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7621 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     7681 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     7741 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
