LOCUS       V043976                 9586 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043976
VERSION     V043976
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             2302..4239
                     /label="HSPA8(NM_006597)"
                     /note="HSPA8(NM_006597)"
                     /gene="HSPA8"
     CDS             4246..4311
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4355..4566
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4579..4847
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4880..5674
                     /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    5681..6269
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6343..6576
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6648..6769
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6788..6923
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6961..6977)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6985..7001
                     /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(7009..7039)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7054..7075
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7363..7951)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8122..8982)
                     /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(8983..9087)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9561..9577
                     /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 agagctagcg aattcgccac catgtccaag ggacctgcag ttggtattga tcttggcacc
     2341 acctactctt gtgtgggtgt tttccagcac ggaaaagtcg agataattgc caatgatcag
     2401 ggaaaccgaa ccactccaag ctatgtcgcc tttacggaca ctgaacggtt gatcggtgat
     2461 gccgcaaaga atcaagttgc aatgaacccc accaacacag tttttgatgc caaacgtctg
     2521 attggacgca gatttgatga tgctgttgtc cagtctgata tgaaacattg gccctttatg
     2581 gtggtgaatg atgctggcag gcccaaggtc caagtagaat acaagggaga gaccaaaagc
     2641 ttctatccag aggaggtgtc ttctatggtt ctgacaaaga tgaaggaaat tgcagaagcc
     2701 taccttggga agactgttac caatgctgtg gtcacagtgc cagcttactt taatgactct
     2761 cagcgtcagg ctaccaaaga tgctggaact attgctggtc tcaatgtact tagaattatt
     2821 aatgagccaa ctgctgctgc tattgcttac ggcttagaca aaaaggttgg agcagaaaga
     2881 aacgtgctca tctttgacct gggaggtggc acttttgatg tgtcaatcct cactattgag
     2941 gatggaatct ttgaggtcaa gtctacagct ggagacaccc acttgggtgg agaagatttt
     3001 gacaaccgaa tggtcaacca ttttattgct gagtttaagc gcaagcataa gaaggacatc
     3061 agtgagaaca agagagctgt aagacgcctc cgtactgctt gtgaacgtgc taagcgtacc
     3121 ctctcttcca gcacccaggc cagtattgag atcgattctc tctatgaagg aatcgacttc
     3181 tatacctcca ttacccgtgc ccgatttgaa gaactgaatg ctgacctgtt ccgtggcacc
     3241 ctggacccag tagagaaagc ccttcgagat gccaaactag acaagtcaca gattcatgat
     3301 attgtcctgg ttggtggttc tactcgtatc cccaagattc agaagcttct ccaagacttc
     3361 ttcaatggaa aagaactgaa taagagcatc aaccctgatg aagctgttgc ttatggtgca
     3421 gctgtccagg cagccatctt gtctggagac aagtctgaga atgttcaaga tttgctgctc
     3481 ttggatgtca ctcctctttc ccttggtatt gaaactgctg gtggagtcat gactgtcctc
     3541 atcaagcgta ataccaccat tcctaccaag cagacacaga ccttcactac ctattctgac
     3601 aaccagcctg gtgtgcttat tcaggtttat gaaggcgagc gtgccatgac aaaggataac
     3661 aacctgcttg gcaagtttga actcacaggc atacctcctg caccccgagg tgttcctcag
     3721 attgaagtca cttttgacat tgatgccaat ggtatactca atgtctctgc tgtggacaag
     3781 agtacgggaa aagagaacaa gattactatc actaatgaca agggccgttt gagcaaggaa
     3841 gacattgaac gtatggtcca ggaagctgag aagtacaaag ctgaagatga gaagcagagg
     3901 gacaaggtgt catccaagaa ttcacttgag tcctatgcct tcaacatgaa agcaactgtt
     3961 gaagatgaga aacttcaagg caagattaac gatgaggaca aacagaagat tctggacaag
     4021 tgtaatgaaa ttatcaactg gcttgataag aatcagactg ctgagaagga agaatttgaa
     4081 catcaacaga aagagctgga gaaagtttgc aaccccatca tcaccaagct gtaccagagt
     4141 gcaggaggca tgccaggagg aatgcctggg ggatttcctg gtggtggagc tcctccctct
     4201 ggtggtgctt cctcagggcc caccattgaa gaggttgatg gatccgacta caaagaccat
     4261 gacggtgatt ataaagatca tgacatcgac tacaaggatg acgatgacaa gtagcggccg
     4321 cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
     4381 cccgagaagt tggggggagg ggtcggcaat tgaacgggtg cctagagaag gtggcgcggg
     4441 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
     4501 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
     4561 acacagctga agcttcgagg ggctcgcatc tctccttcac gcgcccgccg ccctacctga
     4621 ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc ccgcctgtgg tgcctcctga
     4681 actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga gaccgggcct ttgtccggcg
     4741 ctcccttgga gcctacctag actcagccgg ctctccacgc tttgcctgac cctgcttgct
     4801 caactctacg tctttgtttc gttttctgtt ctgcgccgtt acagatccaa gctgtgaccg
     4861 gcgcctacgc taggccacca tggctgaaca agatggattg cacgcaggtt ctccggccgc
     4921 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4981 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5041 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5101 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5161 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5221 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5281 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5341 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5401 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5461 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5521 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5581 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5641 cgccttctat cgccttcttg acgagttctt ctgagtcgac aatcaacctc tggattacaa
     5701 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     5761 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     5821 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     5881 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     5941 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6001 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6061 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     6121 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6181 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6241 tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg acttacaagg
     6301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     6361 cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     6421 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     6481 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     6541 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     6601 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     6661 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6721 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     6781 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6841 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     6901 gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt cgtaatcatg
     6961 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     7021 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     7081 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     7141 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     7201 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     7261 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     7321 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     7381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     7441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     7501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     7561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7801 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     7981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     8041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     8101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     9001 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     9061 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     9121 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     9181 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     9241 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     9301 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     9361 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     9421 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     9481 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     9541 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
