LOCUS       V022817                 9362 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022817
VERSION     V022817
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             683..4042
                     /label="TBC1D8B(NM_017752)"
                     /note="TBC1D8B(NM_017752)"
                     /gene="TBC1D8B"
     CDS             4049..4114
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4138..4726
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4792..4840
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5042..5470
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5484..5813
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5880..6674
                     /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    6850..6971
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7020..7036)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7044..7060
                     /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(7068..7098)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7113..7134
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7422..8010)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8181..9041)
                     /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(9042..9146)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgtggct gaagcctgag gaagtgcttc tgaaaaatgc
      721 gctgaagctg tggctgatgg aaaggtccaa cgactacttc gtgctgcagc ggcgtcgggg
      781 ctacggggag gaaggcggag gggggctcac agggcttctg gttgggactc ttgattcagt
      841 cttggactct actgctaaag tagctccatt tcgcatccta caccagacac cagattctca
      901 agtttacttg tcaattgcat gtggagccaa cagagaagaa ataaccaagc attgggattg
      961 gttggaacaa aatattatga agaccttatc tgtatttgat tcaaatgaag atattactaa
     1021 ttttgtacaa ggaaaaataa gaggattaat tgctgaagag ggaaaacatt gttttgcaaa
     1081 agaagatgat cctgagaaat ttcgagaagc ccttttgaaa tttgaaaaat gttttggttt
     1141 accagagaag gagaagttag tgacctatta ttcatgcagt tattggaaag gacgggttcc
     1201 ttgtcagggt tggctttatc ttagcaccaa ctttctgagc ttctattctt ttttgttggg
     1261 atcagaaata aaactcatta tctcctggga tgaagtctca aaacttgaaa agacttcaaa
     1321 tgtcatactg acagagagta ttcacgtgtg ttcccaagga gagaatcact acttttcaat
     1381 gtttttgcac attaaccaaa cataccttct tatggaacag ctggcaaact atgccattag
     1441 aagacttttt gataaggaaa catttgataa tgacccagtc ctttataatc ctctacagat
     1501 caccaaaaga ggtctggaaa atagagccca cagtgagcaa tttaatgcct tttttaggct
     1561 gcccaaagga gagagtttga aagaagtaca tgaatgtttc ttatgggtac cattcagcca
     1621 cttcaatact catgggaaaa tgtgcatctc agaaaattat atctgctttg ctagccaaga
     1681 tggcaatcag tgtagtgtaa tcattccact acgagaggtc ttagctatag ataagacaaa
     1741 tgattccagc aaatctgtca tcattagcat caaaggaaaa acagcttttc gcttccatga
     1801 agttaaagac tttgaacaac tggtagcaaa actcaggctc agatgcggag cagcttcaac
     1861 tcaatatcat gatattagca cagagcttgc tattagttct gagtctacag agccatctga
     1921 taattttgag gtgcaatctt tgacaagtca gagggaatgc agtaaaactg tgaacactga
     1981 agccttaatg acagtatttc accctcagaa tttggagact cttaattcta aaatgttgaa
     2041 agaaaaaatg aaggaacagt catggaaaat actgtttgca gaatgtggac gtggtgttag
     2101 tatgtttcga accaaaaaga ctcgagatct tgttgtaaga gggattccag aaacattaag
     2161 aggagaactc tggatgcttt tttcaggtgc tgttaatgac atggctacta atcctgacta
     2221 ttatactgaa gtggttgagc agtccttagg gacctgcaac ttggctactg aagaaattga
     2281 acgtgattta cgtcgctctc tgcctgagca cccagccttt cagagtgata ctggcatatc
     2341 tgctctgaga agggtactca cagcttatgc atacaggaat cccaaaattg gatactgcca
     2401 ggcaatgaat attttgactt cagtgctgct tctatatgca aaagaggaag aagctttttg
     2461 gcttctggtt gctgtatgtg aacgaatgtt gcctgattat tttaatcgtc gaattattgg
     2521 tgccttggtg gatcaggcag tctttgaaga acttatcagg gatcaccttc ctcagctgac
     2581 agaacacatg actgatatga cattcttttc ctcagtttct ctctcttggt ttctcacact
     2641 ttttattagt gtgctaccta ttgaaagtgc agtgaatgtg gtggactgtt tcttctatga
     2701 tggaataaag gccattttgc aactgggatt ggcaatactt gactataatt tagacaaact
     2761 gctgacttgt aaagatgatg ctgaagctgt gacagcctta aacaggttct ttgacaatgt
     2821 cactaataag gatagtccat tgccttcaaa tgttcagcaa ggttcaaatg tgagtgatga
     2881 aaaaaccagt catactagag tggatattac agatttgatt agagaatcaa atgagaaata
     2941 tggtaatatt cgctatgaag atatacatag tatgcgctgt cgaaataggt tgtatgtgat
     3001 acagacccta gaggaaacaa caaaacagaa tgtgttgcgt gttgtatcac aagatgtgaa
     3061 attgagcctt caagaattgg atgaacttta tgtcatcttt aagaaagagc tgtttttatc
     3121 ttgttattgg tgtttgggtt gcccagtatt gaagcatcat gaccccagtc tgccatattt
     3181 ggaacagtat cagattgact gccagcagtt cagagcgttg tatcacttgt tgagtccctg
     3241 ggctcattct gcaaataaag actcactagc tttatggaca ttcagattgt tagatgaaaa
     3301 ctctgattgc cttataaact tcaaagaatt ctcctctgca attgacataa tgtacaatgg
     3361 aagttttact gagaagctta agctgctttt taagctacat attcctccag cttacactga
     3421 agtgaaatct aaggatgctt caaaaggaga tgaactttcc aaggaagaat tactttattt
     3481 cagtcagctg catgtttcca agcctgcaaa tgagaaggaa gcagaatcag caaaacacag
     3541 ccctgaaaaa ggcaaaggga aaattgatat tcaagcatat ctaagtcaat ggcaagatga
     3601 gcttttcaaa aaagaagaaa acattaagga tttaccaaga atgaatcagt ctcagtttat
     3661 tcagttttca aagaccctct ataacttatt tcatgaggac cctgaagaag aatcattata
     3721 tcaagccatt gctgttgtaa ccagcctttt actcaggatg gaagaagttg gaaggaaact
     3781 acatagccct acatcatcag ccaaaggatt ctctggtact gtctgtggtt ctggaggacc
     3841 cagtgaggaa aaaacaggga gccacttgga gaaagatcct tgttccttta gggaggaacc
     3901 tcagtggtca tttgcatttg aacagattct tgcatcgctg ttgaatgaac cagcattggt
     3961 gaggtttttt gagaaaccca tagatgtaaa agccaagctg gaaaatgcaa gaatttctca
     4021 gttaaggtct agaaccaaga tgctcgagga ctacaaagac catgacggtg attataaaga
     4081 tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata tctcgacaat
     4141 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4201 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4261 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4321 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4381 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4441 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4501 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     4561 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4621 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4681 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg aaacggggga
     4741 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4801 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4861 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4921 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4981 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
     5041 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5101 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5161 acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg gttccgattt
     5221 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5281 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5341 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5401 taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5461 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     5521 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     5581 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5641 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     5701 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     5761 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     5821 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     5881 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5941 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     6001 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     6061 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     6121 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     6181 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     6241 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     6301 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     6361 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     6421 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     6481 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     6541 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     6601 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     6661 acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac
     6721 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6781 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6841 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6961 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     7021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     7081 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     7141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     7201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     7261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     7321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     7621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7861 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     8041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     8101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     8161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     8221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     8281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     8341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     9001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     9061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     9121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     9181 ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc tctgatgccg
     9241 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     9301 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     9361 gg
//
