LOCUS       V047580                 8987 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047580
VERSION     V047580
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             2296..2730
                     /label="TOMM20(NM_014765)"
                     /note="TOMM20(NM_014765)"
                     /gene="TOMM20"
     CDS             2767..3714
                     /codon_start=1
                     /label="ORF frame 1"
                     /product="bright monomeric near-infrared fluorescent
                     protein engineered from a bacterial phytochrome, with an
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /transl_table=1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES*"
     promoter        3756..3967
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3980..4248
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4281..5075
                     /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="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    5082..5670
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5744..5977
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6049..6170
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6189..6324
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6362..6378)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6386..6402
                     /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(6410..6440)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6455..6476
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6764..7352)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7523..8383)
                     /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(8384..8488)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8962..8978
                     /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 agactcgagg ccaccatggt gggtcggaac agcgccatcg ccgccggtgt atgcggggcc
     2341 cttttcattg ggtactgcat ctacttcgac cgcaaaagac gaagtgaccc caacttcaag
     2401 aacaggcttc gagaacgaag aaagaaacag aagcttgcca aggagagagc tgggctttcc
     2461 aagttacctg accttaaaga tgctgaagct gttcagaagt tcttccttga agaaatacag
     2521 cttggtgaag agttactagc tcaaggtgaa tatgagaagg gcgtagacca tctgacaaat
     2581 gcaattgctg tgtgtggaca gccacagcag ttactgcagg tcttacagca aactcttcca
     2641 ccaccagtgt tccagatgct tctgactaag ctcccaacaa ttagtcagag aattgtaagt
     2701 gctcagagct tggctgaaga tgatgtggaa gaattcggtg gaggcggatc aggaggcggt
     2761 ggatctatgg tagcaggtca tgcctctggc agccccgcat tcgggaccgc ctctcattcg
     2821 aattgcgaac atgaagagat ccacctcgcc ggctcgatcc agccgcatgg cgcgcttctg
     2881 gtcgtcagcg aacatgatca tcgcgtcatc caggccagcg ccaacgccgc ggaatttctg
     2941 aatctcggaa gcgtactcgg cgttccgctc gccgagatcg acggcgatct gttgatcaag
     3001 atcctgccgc atctcgatcc caccgccgaa ggcatgccgg tcgcggtgcg ctgccggatc
     3061 ggcaatccct ctacggagta ctgcggtctg atgcatcggc ctccggaagg cgggctgatc
     3121 atcgaactcg aacgtgccgg cccgtcgatc gatctgtcag gcacgctggc gccggcgctg
     3181 gagcggatcc gcacggcggg ttcactgcgc gcgctgtgcg atgacaccgt gctgctgttt
     3241 cagcagtgca ccggctacga ccgggtgatg gtgtatcgtt tcgatgagca aggccacggc
     3301 ctggtattct ccgagtgcca tgtgcctggg ctcgaatcct atttcggcaa ccgctatccg
     3361 tcgtcgactg tcccgcagat ggcgcggcag ctgtacgtgc ggcagcgcgt ccgcgtgctg
     3421 gtcgacgtca cctatcagcc ggtgccgctg gagccgcggc tgtcgccgct gaccgggcgc
     3481 gatctcgaca tgtcgggctg cttcctgcgc tcgatgtcgc cgtgccatct gcagttcctg
     3541 aaggacatgg gcgtgcgcgc caccctggcg gtgtcgctgg tggtcggcgg caagctgtgg
     3601 ggcctggttg tctgtcacca ttatctgccg cgcttcatcc gtttcgagct gcgggcgatc
     3661 tgcaaacggc tcgccgaaag gatcgcgacg cggatcaccg cgcttgagag ctaagcggcc
     3721 gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
     3781 ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg
     3841 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
     3901 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
     3961 aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg
     4021 aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg
     4081 aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc
     4141 gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc
     4201 tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc
     4261 ggcgcctacg ctaggccacc atggctgaac aagatggatt gcacgcaggt tctccggccg
     4321 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4381 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4441 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4501 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4561 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4621 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4681 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4741 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4801 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4861 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4921 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4981 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5041 tcgccttcta tcgccttctt gacgagttct tctgagtcga caatcaacct ctggattaca
     5101 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     5161 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     5221 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     5281 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     5341 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     5401 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     5461 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga
     5521 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     5581 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     5641 gtcggatctc cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag
     5701 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     5761 tcccaacgaa aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc
     5821 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5881 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5941 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     6001 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt
     6061 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6121 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6181 ctctagctat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     6241 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     6301 agtgaggagg cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat
     6361 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     6421 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     6481 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     6541 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     6601 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     6661 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     6721 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6781 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6841 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6901 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6961 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7021 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7081 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7141 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7201 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7261 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7321 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7381 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7441 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7501 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7561 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7621 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7681 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7741 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7801 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7861 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7921 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7981 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8041 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8101 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     8161 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8221 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8281 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8341 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8521 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8581 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     8641 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     8701 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     8761 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     8821 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     8881 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     8941 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
