LOCUS       V033020                 7935 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033020
VERSION     V033020
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..2615
                     /label="SLC6A6(NM_003043)"
                     /note="SLC6A6(NM_003043)"
                     /gene="SLC6A6"
     CDS             2622..2687
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2711..3299
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3365..3413
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3615..4043
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4057..4386
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4453..5247
                     /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    5423..5544
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5593..5609)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5617..5633
                     /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(5641..5671)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5686..5707
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5995..6583)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6754..7614)
                     /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(7615..7719)
                     /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 caccaaggag aagctgcagt
      781 gtctgaaaga tttccacaag gacatcctga agccctcacc agggaagagc ccaggcacgc
      841 ggcctgagga cgaggctgag ggaaaacctc cgcagaggga gaagtggtct agcaagatcg
      901 actttgtgct ctctgtggct ggcggcttcg tgggcttggg caacgtctgg cgcttcccgt
      961 acctctgcta caagaatggt ggaggtgcgt ttctcatacc gtattttatt ttcctgtttg
     1021 ggagcggcct gcctgtgttt ttcttggaga tcatcatagg ccagtacacc tctgaagggg
     1081 gcatcacctg ctgggaaaag atctgcccct tgttctctgg tatcggctat gcctccgttg
     1141 taattgtgtc cctcctgaat gtctactaca tcgtcatcct ggcctgggcc acatactacc
     1201 tgttccagtc cttccagaag gagctgccct gggcacactg caaccacagc tggaacacac
     1261 ctcactgcat ggaggacacc atgcgcaaga acaagagtgt ctggatcacc atcagctcca
     1321 ccaacttcac ctcccctgtc atcgagttct gggagcgcaa cgtgctgagc ttgtcccctg
     1381 gaatcgacca cccaggctct ctgaaatggg acctcgctct ctgccttctt ttagtctggc
     1441 tagtgtgttt cttctgcatc tggaagggcg tcaggtccac tgggaaggtc gtctacttca
     1501 cagccacttt tccattcgcc atgctcctgg tgctgctggt ccgagggctg acgctgccgg
     1561 gcgcgggcgc aggcatcaag ttctatctgt atcctgacat cacccgcctt gaggacccac
     1621 aggtgtggat tgacgctggg actcagatat tcttctctta tgccatctgc ctgggggcta
     1681 tgacctcgct ggggagctac aacaagtaca agtataactc gtacagggac tgtatgctgc
     1741 tgggatgcct gaacagtggt accagttttg tgtctggctt cgcaattttt tccatcctgg
     1801 gcttcatggc acaagagcaa ggggtggaca ttgctgatgt ggctgagtca ggtcctggcc
     1861 tggccttcat tgcctaccca aaagctgtga caatgatgcc gctgcccaca ttttggtcca
     1921 ttcttttttt tattatgctt ctcttgcttg gactggatag ccagtttgtt gaagttgaag
     1981 gacagatcac atccttggtt gatctttacc catccttcct aaggaagggt tatcgtcggg
     2041 aaatcttcat cgccttcgtg tgtagcatca gctacctgct ggggctgacg atggtgacgg
     2101 agggtggcat gtatgtgttt cagctctttg actactatgc agctagcggt gtatgccttt
     2161 tgtgggttgc attctttgaa tgttttgtta ttgcctggat atatggaggt gataaccttt
     2221 atgatggtat tgaggacatg attggctatc ggcccgggcc ctggatgaag tacagctggg
     2281 ctgtgatcac tccagttctc tgtgttggat gtttcatctt ctcgctcgtc aagtacgtac
     2341 ccctgaccta caacaaaaca tacgtgtacc ccaactgggc cattgggctg ggctggagcc
     2401 tggccctttc ctccatgctc tgcgttccct tggtcatcgt catccgcctc tgccagactg
     2461 aggggccgtt ccttgtgaga gtcaagtacc tgctgacccc aagggaaccc aaccgctggg
     2521 ctgtggagcg cgagggagcc acaccttaca actctcgcac cgtcatgaac ggcgctctcg
     2581 tgaaaccgac ccacatcatt gtggagacca tgatgggatc cgactacaaa gaccatgacg
     2641 gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg
     2701 atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2761 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2821 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2881 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2941 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     3001 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     3061 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     3121 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     3181 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     3241 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     3301 tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     3361 atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg
     3421 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     3481 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     3541 gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc
     3601 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3661 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3721 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt
     3781 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3841 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3901 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3961 ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat
     4021 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4081 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4141 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4201 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4261 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4321 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4381 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4441 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4501 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4561 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4621 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4681 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4741 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4801 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4861 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4921 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4981 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5041 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5101 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5161 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5221 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca
     5281 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5341 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5401 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     5461 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5521 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     5581 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     5641 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5701 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     5761 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5821 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5881 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5941 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6001 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6061 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6121 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6181 gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     6241 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6301 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6361 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6421 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6481 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6541 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6601 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6661 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6721 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6781 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6841 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6901 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6961 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     7021 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     7081 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     7141 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     7201 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     7261 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7321 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7381 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7441 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7501 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7561 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7621 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7681 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7741 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc
     7801 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7861 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7921 aagaatctgc ttagg
//
