LOCUS       V029350                 9534 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029350
VERSION     V029350
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        65..368
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        369..572
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     protein_bind    595..619
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB1"
     CDS             619..2433
                     /label="USP2(NM_004205)"
                     /note="USP2(NM_004205)"
                     /gene="USP2"
     protein_bind    complement(2432..2456)
                     /bound_moiety="BP Clonase™"
                     /gene="mutant version of&nbsp;attB"
                     /note="recombination site for the Gateway® BP reaction"
                     /label="attB2"
     CDS             2458..2499
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     misc_feature    2541..3129
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             3201..3434
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    3512..3633
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      3673..3808
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(3829..3847)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(3857..3873)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      4015..4470
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4496..4600
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             4601..5461
                     /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"
     rep_origin      5632..6220
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    6508..6529
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        6544..6574
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6582..6598
                     /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"
     primer_bind     6606..6622
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        6643..6661
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        6689..6915
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             6916..7096
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    7143..7268
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    7761..7994
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             8179..8223
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     promoter        8398..8898
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     CDS             8910..9305
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
                     /label="BSD"
     misc_feature    9365..9482
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
ORIGIN
        1 cccggggtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctctctggct aactgtcggg atcaacaagt
      601 ttgtacaaaa aagttggcat gtcccagctc tcctccaccc tgaagcgcta cacagaatcg
      661 gcccgctaca cagatgccca ctatgccaag tcgggctatg gtgcctacac cccgtcctcc
      721 tatggggcca atctggctgc ctccttactg gagaaggaga aacttggttt caagccggtc
      781 cccaccagca gcttcctcac ccgtccccgt acctatggcc cctcctccct cctggactat
      841 gaccggggcc gccccctgct gagacccgac atcactgggg gtggtaagcg ggcagagagc
      901 cagacccggg gtactgagcg gcctttaggc agtggcctca gcgggggcag cggattccct
      961 tatggagtga ccaacaactg cctcagctac ctgcccatca atgcctatga ccagggggtg
     1021 accctaaccc agaagctgga cagccaatca gacctggccc gggatttctc cagcctccgg
     1081 acctcagata gctaccggat agaccccagg aacctgggcc gcagccccat gctggcccgg
     1141 acgcgcaagg agctctgcac cctgcagggg ctctaccaga cagccagctg ccctgaatac
     1201 ctggtcgact acctggagaa ctatggtcgc aagggcagtg catctcaggt gccctcccag
     1261 gcccctccct cacgagtccc tgaaatcatc agcccaacct accgacccat tggccgctac
     1321 acgctgtggg agacgggaaa gggtcaggcc cctgggccca gccgctccag ctccccggga
     1381 agagacggca tgaattctaa gagtgcccag ggtctggctg gtcttcgaaa ccttgggaac
     1441 acgtgcttca tgaactcaat tctgcagtgc ctgagcaaca ctcgggagtt gagagattac
     1501 tgcctccaga ggctctacat gcgggacctg caccacggca gcaatgcaca cacagccctc
     1561 gtggaagagt ttgcaaaact aattcagacc atatggactt catcccccaa tgatgtggtg
     1621 agcccatctg agttcaagac ccagatccag agatacgcac cgcgctttgt tggctataat
     1681 cagcaggatg ctcaggagtt ccttcgcttt cttctggatg ggctccataa cgaggtgaac
     1741 cgagtgacac tgagacctaa gtccaaccct gagaacctcg atcatcttcc tgatgacgag
     1801 aaaggccgac agatgtggag aaaatatcta gaacgggaag acagtaggat cggggatctc
     1861 tttgttgggc agctaaagag ctcgctgacg tgtacagatt gtggttactg ttctacggtc
     1921 ttcgacccct tctgggacct ctcactgccc attgctaagc gaggttatcc tgaggtgaca
     1981 ttaatggact gcatgaggct cttcaccaaa gaggatgtgc ttgatggaga tgaaaagcca
     2041 acatgctgtc gctgccgagg cagaaaacgg tgtataaaga agttctccat ccagaggttc
     2101 ccaaagatct tggtgctcca tctgaagcgg ttctcagaat ccaggatccg aaccagcaag
     2161 ctcacaacat ttgtgaactt ccccctaaga gacctggact taagagaatt tgcctcagaa
     2221 aacaccaacc atgctgttta caacctgtac gctgtgtcca atcactccgg aaccaccatg
     2281 ggtggccact atacagccta ctgtcgcagt ccagggacag gagaatggca cactttcaac
     2341 gactccagcg tcactcccat gtcctccagc caagtgcgca ccagcgacgc ctacctgctc
     2401 ttctacgaac tggccagccc gccctcccga atgccaactt tcttgtacaa agtggttggt
     2461 aagcctatcc ctaaccctct cctcggtctc gattctacgt agtaatgagc tagcgctaac
     2521 cggtggcgcg ttaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2581 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2641 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2701 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2761 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2821 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2881 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     2941 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3001 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3061 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     3121 cctccccgcg tcgactttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     3181 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     3241 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     3301 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     3361 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     3421 gaaaatctct agcagtacgt atagtagttc atgtcatctt attattcagt atttataact
     3481 tgcaaagaaa tgaatatcag agagtgagag gaacttgttt attgcagctt ataatggtta
     3541 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3601 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctctagc tatcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta gggacgtacc caattcgccc tatagtgagt
     3841 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     3901 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     3961 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     4021 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4081 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4141 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4201 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4261 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4321 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4381 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4441 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     4501 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4561 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     4621 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4681 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     4741 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     4801 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4861 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     4921 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4981 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     5041 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5101 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5161 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5221 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5281 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5341 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5401 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5461 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     5521 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     5581 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     5641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     5701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     5761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     5821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     5881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     5941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     6001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     6061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     6121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     6181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     6241 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     6301 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6361 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     6421 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6481 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     6541 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     6601 tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc ctcactaaag
     6661 ggaacaaaag ctggagctgc aagcttaatg tagtcttatg caatactctt gtagtcttgc
     6721 aacatggtaa cgatgagtta gcaacatgcc ttacaaggag agaaaaagca ccgtgcatgc
     6781 cgattggtgg aagtaaggtg gtacgatcgt gccttattag gaaggcaaca gacgggtctg
     6841 acatggattg gacgaaccac tgaattgccg cattgcagag atattgtatt taagtgccta
     6901 gctcgataca taaacgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6961 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7021 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7081 tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa agcgaaaggg aaaccagagg
     7141 agctctctcg acgcaggact cggcttgctg aagcgcgcac ggcaagaggc gaggggcggc
     7201 gactggtgag tacgccaaaa attttgacta gcggaggcta gaaggagaga gatgggtgcg
     7261 agagcgtcag tattaagcgg gggagaatta gatcgcgatg ggaaaaaatt cggttaaggc
     7321 cagggggaaa gaaaaaatat aaattaaaac atatagtatg ggcaagcagg gagctagaac
     7381 gattcgcagt taatcctggc ctgttagaaa catcagaagg ctgtagacaa atactgggac
     7441 agctacaacc atcccttcag acaggatcag aagaacttag atcattatat aatacagtag
     7501 caaccctcta ttgtgtgcat caaaggatag agataaaaga caccaaggaa gctttagaca
     7561 agatagagga agagcaaaac aaaagtaaga ccaccgcaca gcaagcggcc gctgatcttc
     7621 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     7681 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     7741 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     7801 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     7861 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     7921 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     7981 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     8041 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     8101 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     8161 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     8221 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     8281 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     8341 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccttg
     8401 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
     8461 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
     8521 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
     8581 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
     8641 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
     8701 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
     8761 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
     8821 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
     8881 accgacctct ctccccaggg ggtaccacca tggccaagcc tttgtctcaa gaagaatcca
     8941 ccctcattga aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg
     9001 tcgccagcgc agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt
     9061 ttactggggg accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg
     9121 gcaacctgac ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg
     9181 gacggtgccg acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca
     9241 gtgatggaca gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg
     9301 agggcctgca gctgcagtag taagaattct agatcttgag acaaatggca gtattcatcc
     9361 acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     9421 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     9481 ttcgggttta ttacagggac agcagagatc cactttggcg ccggctcgag gggg
//
