LOCUS       V042792                 9964 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042792
VERSION     V042792
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..4818
                     /label="ATG9A(NM_024085)"
                     /note="ATG9A(NM_024085)"
                     /gene="ATG9A"
     CDS             4825..4890
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4934..5145
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5158..5426
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5453..6052
                     /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    6059..6647
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6721..6954
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7026..7147
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7166..7301
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7339..7355)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7363..7379
                     /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(7387..7417)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7432..7453
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7741..8329)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8500..9360)
                     /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(9361..9465)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9939..9955
                     /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 catggcgcag tttgacactg aataccagcg cctagaggcc
     2341 tcctatagtg attcaccccc aggggaggag gacctgttgg tgcacgtcgc cgaggggagc
     2401 aagtcacctt ggcaccatat tgaaaacctt gacctcttct tctctcgagt ttataatctg
     2461 caccagaaga atggcttcac atgtatgctc atcggggaga tctttgagct catgcagttc
     2521 ctctttgtgg ttgccttcac taccttcctg gtcagctgcg tggactatga catcctattt
     2581 gccaacaaga tggtgaacca cagtcttcac cctactgaac ccgtcaaggt cactctgcca
     2641 gacgcctttt tgcctgctca agtctgtagt gccaggattc aggaaaatgg ctcccttatc
     2701 accatcctgg tcattgctgg tgtcttctgg atccaccggc ttatcaagtt catctataac
     2761 atttgctgct actgggagat ccactccttc tacctgcacg ctctgcgcat ccctatgtct
     2821 gcccttccgt attgcacgtg gcaagaagtg caggcccgga tcgtgcagac gcagaaggag
     2881 caccagatct gcatccacaa acgtgagctg acagaactgg acatctacca ccgcatcctc
     2941 cgtttccaga actacatggt ggcactggtt aacaaatccc tcctgcctct gcgcttccgc
     3001 ctgcctggcc tcggggaagc tgtcttcttc acccgtggtc tcaagtacaa ctttgagctg
     3061 atcctcttct ggggacctgg ctctctgttt ctcaatgaat ggagcctcaa ggccgagtac
     3121 aaacgtgggg ggcaacggct agagctggcc cagcgcctca gcaaccgcat cctgtggatt
     3181 ggcatcgcta acttcctgct gtgccccctc atcctcatat ggcaaatcct ctatgccttc
     3241 ttcagctatg ctgaggtgct gaagcgggag ccgggggccc tgggagcacg ctgctggtca
     3301 ctctatggcc gctgctacct ccgccacttc aacgagctgg agcacgagct gcagtcccgc
     3361 ctcaaccgtg gctacaagcc cgcctccaag tacatgaatt gcttcttgtc acctcttttg
     3421 acactgctgg ccaagaatgg agccttcttc gctggctcca tcctggctgt gcttattgcc
     3481 ctcaccattt atgacgaaga tgtgttggct gtggaacatg tgctgaccac cgtcacactc
     3541 ctgggggtca ccgtgaccgt gtgcaggtcc tttatcccgg accagcacat ggtgttctgc
     3601 cctgagcagc tgctccgcgt gatcctcgct cacatccact acatgcctga ccactggcag
     3661 ggtaatgccc accgctcgca gacccgggac gagtttgccc agctcttcca gtacaaggca
     3721 gtgttcattt tggaagagtt gctgagcccc attgtcacac ccctcatcct catcttctgc
     3781 ctgcgcccac gggccctgga gattatagac ttcttccgaa acttcaccgt ggaggtcgtt
     3841 ggtgtgggag atacctgctc ctttgctcag atggatgttc gccagcatgg tcatccccag
     3901 tggctatctg ctgggcagac agaggcctca gtgtaccagc aagctgagga tggaaagaca
     3961 gagttgtcac tcatgcactt tgccatcacc aaccctggct ggcagccacc acgtgagagc
     4021 acagccttcc taggcttcct caaggagcag gttcagcggg atggagcagc tgctagcctc
     4081 gcccaagggg gtctgctccc tgaaaatgcc ctctttacgt ctatccagtc cttacaatct
     4141 gagtctgagc ccctgagcct tatcgcaaat gtggtagctg gctcatcctg ccggggccct
     4201 ccactgccca gagacctgca gggctccagg cacagggctg aagtcgcctc tgccctgcgc
     4261 tccttctccc cgctgcaacc cgggcaggcg cccacaggcc gggctcacag caccatgaca
     4321 ggctctgggg tggatgccag gacagccagc tccgggagca gcgtgtggga aggacagctg
     4381 cagagcctgg tgctgtcaga atatgcatcc acagagatga gcctgcatgc cctctatatg
     4441 caccagctcc acaagcagca ggcccaggct gaacctgagc ggcatgtatg gcaccgccgg
     4501 gagagtgatg agagtggaga aagcgcccct gatgaagggg gagagggcgc ccgggccccc
     4561 cagtctatcc ctcgctctgc tagctatccc tgtgcagcac cccggcctgg agctcctgag
     4621 accactgccc tgcatggggg cttccagagg cgctacggtg gcatcacaga tcctggcaca
     4681 gtgcccaggg ttccctctca tttctctcgg ctgcctcttg gagggtgggc agaagatggg
     4741 cagtcggcat caaggcaccc tgagcccgtg cccgaagagg gctcggagga tgagctaccc
     4801 cctcaggtgc acaaggtagg atccgactac aaagaccatg acggtgatta taaagatcat
     4861 gacatcgact acaaggatga cgatgacaag tagcggccgc gaaggatctg cgatcgctcc
     4921 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg
     4981 gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc
     5041 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc
     5101 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa gcttcgaggg
     5161 gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg
     5221 agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta
     5281 agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag cctacctaga
     5341 ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg
     5401 ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct agatgaccga
     5461 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct
     5521 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat
     5581 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     5641 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     5701 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     5761 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     5821 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     5881 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     5941 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     6001 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gagtcgacaa
     6061 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6121 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     6181 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6241 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6301 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6361 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6421 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
     6481 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6541 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6601 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa
     6661 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6721 tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt actgggtctc
     6781 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     6841 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     6901 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     6961 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     7021 agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     7081 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     7141 tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt ccgcccattc
     7201 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     7261 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt gcagagacgg
     7321 cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     7381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     7441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     7501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     7561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     7621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     8041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     8101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     8161 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     8221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     8281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     8341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     8401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     8461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     8521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     8581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     8641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     8701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     8761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     8821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     8881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     8941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     9001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     9061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     9121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     9181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     9241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     9301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     9361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     9421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     9481 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     9541 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     9601 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     9661 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     9721 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     9781 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     9841 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     9901 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
     9961 gctg
//
