LOCUS       V046689                 9762 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046689
VERSION     V046689
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..4442
                     /label="Aff3(NM_001290814)"
                     /note="Aff3(NM_001290814)"
                     /gene="Aff3"
     CDS             4449..4514
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4538..5126
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5192..5240
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5442..5870
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5884..6213
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6280..7074
                     /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    7250..7371
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7420..7436)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7444..7460
                     /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(7468..7498)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7513..7534
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7822..8410)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8581..9441)
                     /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(9442..9546)
                     /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 ccaccatgga cagcttcgac ttagccctgc
      781 tccaggaatg ggacctcgag tcgctgtgtg tctatgaacc ggacaggaat gcgttacgga
      841 ggaaagagcg agaaagaaga agtcaagaaa cacagcagga cagtggcagc tttaactccg
      901 gttactcgct cttcagcgaa ccctacaaga cgaacaaggg ggatgaactc tccaaccgga
      961 tccagaatac attaggaaat tatgatgaga tgaaagactt tttaactgat agatccaacc
     1021 agagtcatct tgttggcgtc cccaagccag gtgtccctca gactcccgtg aacaagatag
     1081 atgagcattt tggggcagag tcgagagccc agccccagcc ctcaaccgtc tgcagcactg
     1141 cgtcctctac acctgcagct gtccctgtgc agcagggcaa gagaggtgcc atgggctggc
     1201 agaaggctgg gcacccaccg tccgatggcc aacagagagc agcgcaacag ggctctctca
     1261 ggaccttgct tggagatggt gttggcagac agcagacaag ggccaagcaa gtgtgcaata
     1321 tggagacggg gcttcagacc caggagaggc cacctgcaat ggcagccaag cacggcggca
     1381 gtggacactg tgttcagaac tttcccccat ccctggcttc aaaacccagc ctggtgcagc
     1441 agaaaccaac tgcctacgtg cggccaatgg atggccagga ccaagctcct gacgaatctc
     1501 ctaagctgaa gtcatccaca gaaactgctg tgcactgcac agcctacaga ggggtcccgg
     1561 ccaacaaacc agagtcagcc cgagccaagg ccaagcttgc caagttcagc atccccaagc
     1621 aggcagagga gagtagatct ggagaaaaca acagctgtgt ggaagaaata attcgggaga
     1681 tgacctggct tcctccactt tctgctattc aagctcctgc caaagtggaa ccaagcaagt
     1741 tcccattccc aaataaggac tcgcaacttg tatcctctgg acacagtaat ccaaagaaag
     1801 ctgatgcgga gccagggagt ccagacaatg gcgcatcgaa cacttcaact ctggaagacg
     1861 accttaagct aagcagtgat gacgaagagg gtgagcagca ggcagcccag agaacagctc
     1921 tccgcgctct agctgacagc tccgtggttc agcagaccaa ctgtagaggt tcggcgccat
     1981 ccagcaaggg cggcggcagc agcagcagca gcggcggcag cagctcctcc agcgactcag
     2041 agagcacttc cggctccgac tccgagacgg agagcagctc cagctccagc gagagcgagg
     2101 gcagcaagcc tccgcactgc tccagccccg aggccgaacc tgcgtcctct aacaagtggc
     2161 agctggataa atggctaaac aaagttaatc ctcacaagcc gccaattctg atccaaaatg
     2221 aaagccacgg cccggaaagg aatcaatact acacgccgcc ggtgaaagat gaagggcaag
     2281 actgcgggaa acttcctgag atctgccaag ccagcctgag ggacaaggag ctcaagacca
     2341 cctgcaagga ggagcagagg ccgaggaccg caaacaaggc ccctggcagc aaaagtgtaa
     2401 agcagaagtc cccacccgct gctgtggctg tgaccgcagc tgccctcccg cccgcggtgc
     2461 ccagcgcacc cacggagagc gcgcctgcgc ccacccgcag gtcagcaggc aagaagccca
     2521 cccggcgcac cgagaggacc tcagccgggg acggtgccaa ctgccaccgg cctgaggagc
     2581 cagtggcccc agacacgctg ggggcgagcg tggtgggacc cctagagccc cccaaaacca
     2641 gaccctgcgg taacaacagg acaggccacc gcaaggagct gcgctcctcc gtgacctgtg
     2701 agaagcggcg cacgaggggg ctgagcagaa tcgtccccaa atccaaggag tttattgaaa
     2761 cagaatcttc gtcttcctct tcctcctcgg actcagacct ggagtcagag caagaggagt
     2821 acgtcctgtc caaggctccg accaccactg ggagcgagca gcggctgaag gaggccgcca
     2881 gcagcaacaa caacagcaac agcaacagca gcacctcccg ggcttctgtg ggctccatca
     2941 acgccaggac caccagtgac attgccaagg agctagagga acagttctac acgctggtcc
     3001 cctttggcag gaatgaacta ctatctccgc taaaggacag cgatgaggtc aggtctctct
     3061 gggtcaaaat tgacctgacc ctcctgtcca ggatcccaga acacctgtcc caggagcctg
     3121 gggttttgag tgctccttcg gccaaggaca ctgatagtgc cccagccagt catgcattag
     3181 acgctcctgc tgaaaagact ttgccaaaat ccaagaggaa acgcaagtgc gacaacgaag
     3241 atgactacag ggagataaag aaggtccagg gaaggaagga gagtgcctca cgcctcgccg
     3301 cctccactaa taacaccttg tccggcaatc actgcaatgt gaacgtcaac agcttggcaa
     3361 taccaataaa caaaaatgaa aagatgctcc ggtcgcccac ttcgccactc tcagacacat
     3421 gtaaacacaa atacgccagc gaggacctga cttcttccag ccgacctcac ggcaacgggt
     3481 tgttgacttc agcgtcttcc aacaaagagc caaaggctga gagtcagctg cagacgattg
     3541 ctggggacct cacgaaggca tctcacaaca gttctgaaaa tggcaccctc catagcaagt
     3601 cacggcccca gacggagcca tggtctccag gttccaatgg ccacagggac tgcaagagac
     3661 agaaactgat cttcgatgac atgcctcgaa gtgcagatta ttttatgcga gaagccaagc
     3721 ggatgaaaca taaagcagac gccatggtgg agaagttcgg gaaggccctg aactatgccg
     3781 aagctgcctt gtccttcatc gagtgcggaa acgcaatgga acaagggcca atggaatcca
     3841 agtctcccta caccatgtac tcggagacag tggagctcat caggtatgct atgagactaa
     3901 aaacccactc aggccccaat gccacaccag aggacaaaca gttagctgcg ctttgttacc
     3961 gatgcctggc tctcctatac tggcggatgt ttcgactcaa aagagaccat gctgtaaaat
     4021 attcaaaagc acttatcgac tatttcaaga attcatcgaa agctgcccaa gccccatccc
     4081 catgggggtc cagtggcaag agcactggaa gcccatcccc catgtctccc aacccttctc
     4141 ccgccagctc cgtggggtct cagggcagcc tctccagctc cagcggcctg tccccatcga
     4201 ccatcgtcag catcccccag cgcattcacc agatggcggc caatcacgtc agcatcacca
     4261 acagcatcct ccacagctat gactactggg agatggctga taacctggcc aaggaaaaca
     4321 gagagttctt caatgacctg gatttgctca tgggacctgt caccctgcac agcagcatgg
     4381 aacacctggt ccagtactcc cagcaaggcc tgcattggct tcggagcagc gtacacctgt
     4441 ccggatccga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     4501 atgacgatga caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat
     4561 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     4621 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     4681 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     4741 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     4801 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     4861 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4921 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
     4981 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5041 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5101 gatctccctt tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag
     5161 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     5221 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5281 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5341 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5401 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5461 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5521 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5581 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     5641 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5701 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5761 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     5821 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5881 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5941 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     6001 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     6061 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6121 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     6181 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     6241 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6301 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6361 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6421 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6481 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6541 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6601 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6661 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6721 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6781 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6841 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6901 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6961 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7021 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     7081 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     7141 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     7201 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     7261 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7321 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7381 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7441 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7501 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7561 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7621 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7681 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7741 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7801 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7861 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7921 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7981 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     8041 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     8101 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     8161 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     8221 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     8281 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8341 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8401 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8461 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8521 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8581 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8641 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8701 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8761 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8821 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8881 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8941 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     9001 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     9061 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     9121 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     9181 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     9241 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9301 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9361 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9421 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9481 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9541 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     9601 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     9661 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     9721 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gg
//
