LOCUS       V034972                 9308 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034972
VERSION     V034972
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             2305..2322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             2329..4194
                     /label="F2(NM_000506)"
                     /note="F2(NM_000506)"
                     /gene="F2"
     CDS             4201..4227
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     promoter        4278..4489
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4502..4770
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4797..5396
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5403..5991
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6065..6298
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6370..6491
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6510..6645
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6683..6699)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6707..6723
                     /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(6731..6761)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6776..6797
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7085..7673)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7844..8704)
                     /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(8705..8809)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9283..9299
                     /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 catgcaccat catcatcatc atgatatcat ggcgcacgtc
     2341 cgaggcttgc agctgcctgg ctgcctggcc ctggctgccc tgtgtagcct tgtgcacagc
     2401 cagcatgtgt tcctggctcc tcagcaagca cggtcgctgc tccagcgggt ccggcgagcc
     2461 aacaccttct tggaggaggt gcgcaagggc aacctggagc gagagtgcgt ggaggagacg
     2521 tgcagctacg aggaggcctt cgaggctctg gagtcctcca cggctacgga tgtgttctgg
     2581 gccaagtaca cagcttgtga gacagcgagg acgcctcgag ataagcttgc tgcatgtctg
     2641 gaaggtaact gtgctgaggg tctgggtacg aactaccgag ggcatgtgaa catcacccgg
     2701 tcaggcattg agtgccagct atggaggagt cgctacccac ataagcctga aatcaactcc
     2761 actacccatc ctggggccga cctacaggag aatttctgcc gcaaccccga cagcagcacc
     2821 acgggaccct ggtgctacac tacagacccc accgtgagga ggcaggaatg cagcatccct
     2881 gtctgtggcc aggatcaagt cactgtagcg atgactccac gctccgaagg ctccagtgtg
     2941 aatctgtcac ctccattgga gcagtgtgtc cctgatcggg ggcagcagta ccaggggcgc
     3001 ctggcggtga ccacacatgg gctcccctgc ctggcctggg ccagcgcaca ggccaaggcc
     3061 ctgagcaagc accaggactt caactcagct gtgcagctgg tggagaactt ctgccgcaac
     3121 ccagacgggg atgaggaggg cgtgtggtgc tatgtggccg ggaagcctgg cgactttggg
     3181 tactgcgacc tcaactattg tgaggaggcc gtggaggagg agacaggaga tgggctggat
     3241 gaggactcag acagggccat cgaagggcgt accgccacca gtgagtacca gactttcttc
     3301 aatccgagga cctttggctc gggagaggca gactgtgggc tgcgacctct gttcgagaag
     3361 aagtcgctgg aggacaaaac cgaaagagag ctcctggaat cctacatcga cgggcgcatt
     3421 gtggagggct cggatgcaga gatcggcatg tcaccttggc aggtgatgct tttccggaag
     3481 agtccccagg agctgctgtg tggggccagc ctcatcagtg accgctgggt cctcaccgcc
     3541 gcccactgcc tcctgtaccc gccctgggac aagaacttca ccgagaatga ccttctggtg
     3601 cgcattggca agcactcccg caccaggtac gagcgaaaca ttgaaaagat atccatgttg
     3661 gaaaagatct acatccaccc caggtacaac tggcgggaga acctggaccg ggacattgcc
     3721 ctgatgaagc tgaagaagcc tgttgccttc agtgactaca ttcaccctgt gtgtctgccc
     3781 gacagggaga cggcagccag cttgctccag gctggataca aggggcgggt gacaggctgg
     3841 ggcaacctga aggagacgtg gacagccaac gttggtaagg ggcagcccag tgtcctgcag
     3901 gtggtgaacc tgcccattgt ggagcggccg gtctgcaagg actccacccg gatccgcatc
     3961 actgacaaca tgttctgtgc tggttacaag cctgatgaag ggaaacgagg ggatgcctgt
     4021 gaaggtgaca gtgggggacc ctttgtcatg aagagcccct ttaacaaccg ctggtatcaa
     4081 atgggcatcg tctcatgggg tgaaggctgt gaccgggatg ggaaatatgg cttctacaca
     4141 catgtgttcc gcctgaagaa gtggatacag aaggtcattg atcagtttgg agagctcgag
     4201 tacccatacg acgtaccaga ttacgcttag ggatccgcgg ccgcgaagga tctgcgatcg
     4261 ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg
     4321 aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga
     4381 tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt
     4441 agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg
     4501 aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg
     4561 gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta
     4621 ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc
     4681 tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt
     4741 ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagatga
     4801 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca
     4861 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc
     4921 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4981 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     5041 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     5101 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5161 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5221 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5281 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5341 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagtcg
     5401 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     5461 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     5521 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     5581 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     5641 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     5701 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     5761 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     5821 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     5881 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     5941 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct
     6001 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     6061 ggactggaag ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg
     6121 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6181 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6241 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6301 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6361 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6481 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc
     6541 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     6601 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag
     6661 acggcccaaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6721 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6781 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6841 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     7021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     7081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     7141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     7201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     7261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     7321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     7381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     7441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7501 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7801 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7861 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7921 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7981 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     8041 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     8101 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     8161 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     8221 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     8281 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     8341 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     8401 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     8461 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8521 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8581 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8641 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8701 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8761 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8821 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     8881 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     8941 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     9001 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     9061 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     9121 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     9181 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     9241 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     9301 ccaagctg
//
