LOCUS       V034754                 9686 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034754
VERSION     V034754
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /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"
     misc_feature    1804..1920
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2010..2213
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2290..2985
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric far-red fluorescent protein (Shcherbo
                     et al., 2009)"
                     /transl_table=1
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
                     /label="mKate2"
     CDS             3022..4161
                     /label="Dek(NM_025900)"
                     /note="Dek(NM_025900)"
                     /gene="Dek"
     CDS             4162..4179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     promoter        4229..4440
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4453..4721
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4749..5774
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /note="confers resistance to hygromycin"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /transl_table=1
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E*"
                     /label="HygR"
     misc_feature    5781..6369
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6443..6676
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6748..6869
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6888..7023
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7061..7077)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7085..7101
                     /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(7109..7139)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7154..7175
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7463..8051)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8222..9082)
                     /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(9083..9187)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9661..9677
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggttta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagccacca tggtgagcga gctgattaag gagaacatgc acatgaagct gtacatggag
     2341 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
     2401 ggcacccaga ccatgagaat caaggcggtc gagggcggcc ctctcccctt cgccttcgac
     2461 atcctggcta ccagcttcat gtacggcagc aaaaccttca tcaaccacac ccagggcatc
     2521 cccgacttct ttaagcagtc cttccccgag ggcttcacat gggagagagt caccacatac
     2581 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
     2641 tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
     2701 acactcggct gggaggcctc caccgagacc ctgtaccccg ctgacggcgg cctggaaggc
     2761 agagccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttgaagacc
     2821 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac
     2881 agaagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg
     2941 gctgtggcca gatactgcga cctccctagc aaactggggc acagaggtgg aggcggatca
     3001 ggaggcggtg gatcttctag aatgtcggcg gcggcggccc ccgctgcgga gggagaggac
     3061 gcccccgtgc cgccctcatc cgagaaggaa cccgagatgc cgggtcccag ggaagagagt
     3121 gaggaggagg aggaggatga cgaagacgat gatgaagagg acgaggagga agaaaaagaa
     3181 aagagtctta tcgtggaagg caagagagag aagaagaaag tagagagact gacgatgcaa
     3241 gtgtcttcct tacagagaga gccatttaca gtgacacaag ggaagggtca gaaactttgt
     3301 gaaattgaaa ggatacattt ctttctgagt aagaaaaaac cagatgaact tagaaatcta
     3361 cacaaactgc tttacaacag gccgggcaca gtgtcctcgt tgaagaagaa cgtgggtcag
     3421 ttcagtggct ttccattcga aaaaggcagt acccagtata aaaagaagga agaaatgttg
     3481 aaaaagtttc gaaatgccat gttaaagagc atctgtgagg ttcttgattt agagaggtca
     3541 ggcgtgaaca gcgaactcgt gaagaggatc ttgaacttct taatgcatcc aaagccttct
     3601 ggcaaaccat taccaaaatc caaaaaatct tccagcaaag gtagtaaaaa ggaacggaac
     3661 agttctggaa caacaaggaa gtcaaagcaa actaaatgcc ctgaaattct gtcagatgag
     3721 tctagtagtg atgaagatga gaagaaaaat aaggaagagt cttcggaaga tgaagagaaa
     3781 gaaagtgaag aggagcaacc accaaaaaag acatctaaaa aagaaaaagc aaaacagaaa
     3841 gctactgcta aaagtaaaaa atctgtgaag agtgctaatg ttaagaaggc agacagcagt
     3901 accaccaaga agaatcaaaa aagttccaaa aaagagtctg aatctgaaga cagttctgat
     3961 gatgaaccct taattaaaaa attgaaaaag ccacctacag atgaagagct aaaggaaaca
     4021 gtgaagaaat tactggctga tgctaacttg gaagaagtca caatgaagca gatttgcaaa
     4081 gaggtatatg aaaattatcc tgcttatgat ttgactgaga ggaaagattt cattaaaaca
     4141 actgtaaaag agctaatttc tcatcaccat catcaccatt gaggatccgc ggccgcaagg
     4201 atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     4261 aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac
     4321 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     4381 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     4441 ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc
     4501 catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg
     4561 tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct
     4621 tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc
     4681 tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct
     4741 acgtcgacat gaaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     4801 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     4861 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     4921 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     4981 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     5041 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     5101 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     5161 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     5221 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     5281 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     5341 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     5401 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     5461 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     5521 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     5581 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     5641 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     5701 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     5761 gggcaaagga ataggtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     5821 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     5881 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     5941 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     6001 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     6061 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     6121 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     6181 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     6241 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     6301 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     6361 cctccccgcc tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6421 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg
     6481 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     6541 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     6601 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     6661 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     6721 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     6781 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6841 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     6901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6961 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7021 gcctagactt ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt
     7081 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7141 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7201 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7261 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7321 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7381 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7441 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     7561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     7621 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     7681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     7741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     7801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     7861 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     7921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     7981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8221 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8281 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8341 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8401 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8461 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     8521 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     8581 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     8641 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     8701 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     8761 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     8821 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     8881 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     8941 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9001 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9061 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9121 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9181 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     9241 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     9301 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     9361 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     9421 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
     9481 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
     9541 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     9601 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     9661 gtaaaacgac ggccagtgcc aagctg
//
