LOCUS       V042017                 9754 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042017
VERSION     V042017
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..4608
                     /label="ITGB8(NM_002214)"
                     /note="ITGB8(NM_002214)"
                     /gene="ITGB8"
     CDS             4615..4680
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4724..4935
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4948..5216
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5243..5842
                     /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    5849..6437
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6511..6744
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6816..6937
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6956..7091
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7129..7145)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7153..7169
                     /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(7177..7207)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7222..7243
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7531..8119)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8290..9150)
                     /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(9151..9255)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9729..9745
                     /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 catgtgcggc tcggccctgg ctttttttac cgctgcattt
     2341 gtctgcctgc aaaacgaccg gcgaggtccc gcctcgttcc tctgggcagc ctgggtgttt
     2401 tcacttgttc ttggactggg ccaaggtgaa gacaatagat gtgcatcttc aaatgcagca
     2461 tcctgtgcca ggtgccttgc gctgggtcca gaatgtggat ggtgtgttca agaggatttc
     2521 atttcaggtg gatcaagaag tgaacgttgt gatattgttt ccaatttaat aagcaaaggc
     2581 tgctcagttg attcaataga atacccatct gtgcatgtta taatacccac tgaaaatgaa
     2641 attaataccc aggtgacacc aggagaagtg tctatccagc tgcgtccagg agccgaagct
     2701 aattttatgc tgaaagttca tcctctgaag aaatatcctg tggatcttta ttatcttgtt
     2761 gatgtctcag catcaatgca caataatata gaaaaattaa attccgttgg aaacgattta
     2821 tctagaaaaa tggcattttt ctcccgtgac tttcgtcttg gatttggctc atacgttgat
     2881 aaaacagttt caccatacat tagcatccac cccgaaagga ttcataatca atgcagtgac
     2941 tacaatttag actgcatgcc tccccatgga tacatccatg tgctgtcttt gacagagaac
     3001 atcactgagt ttgagaaagc agttcataga cagaagatct ctggaaacat agatacacca
     3061 gaaggaggtt ttgacgccat gcttcaggca gctgtctgtg aaagtcatat cggatggcga
     3121 aaagaggcta aaagattgct gctggtgatg acagatcaga cgtctcatct cgctcttgat
     3181 agcaaattgg caggcatagt ggtgcccaat gacggaaact gtcatctgaa aaacaacgtc
     3241 tatgtcaaat cgacaaccat ggaacacccc tcactaggcc aactttcaga gaaattaata
     3301 gacaacaaca ttaatgtcat ctttgcagtt caaggaaaac aatttcattg gtataaggat
     3361 cttctacccc tcttgccagg caccattgct ggtgaaatag aatcaaaggc tgcaaacctc
     3421 aataatttgg tagtggaagc ctatcagaag ctcatttcag aagtgaaagt tcaggtggaa
     3481 aaccaggtac aaggcatcta ttttaacatt accgccatct gtccagatgg gtccagaaag
     3541 ccaggcatgg aaggatgcag aaacgtgacg agcaatgatg aagttctttt caatgtaaca
     3601 gttacaatga aaaaatgtga tgtcacagga ggaaaaaact atgcaataat caaacctatt
     3661 ggttttaatg aaaccgctaa aattcatata cacagaaact gcagctgtca gtgtgaggac
     3721 aacagaggac ctaaaggaaa gtgtgtagat gaaacttttc tagattccaa gtgtttccag
     3781 tgtgatgaga ataaatgtca ttttgatgaa gatcagtttt cttctgagag ttgcaagtca
     3841 cacaaggatc agcctgtttg cagtggtcga ggagtttgtg tttgtgggaa atgttcatgt
     3901 cacaaaatta agcttggaaa agtgtatgga aaatactgtg aaaaggatga cttttcttgt
     3961 ccatatcacc atggaaatct gtgtgctggg catggagagt gtgaagcagg cagatgccaa
     4021 tgcttcagtg gctgggaagg tgatcgatgc cagtgccctt cagcagcagc ccagcactgt
     4081 gtcaattcaa agggccaagt gtgcagtgga agaggcacgt gtgtgtgtgg aaggtgtgag
     4141 tgcaccgatc ccaggagcat cggccgcttc tgtgaacact gccccacctg ttatacagcc
     4201 tgcaaggaaa actggaattg tatgcaatgc cttcaccctc acaatttgtc tcaggctata
     4261 cttgatcagt gcaaaacctc atgtgctctc atggaacaac agcattatgt cgaccaaact
     4321 tcagaatgtt tctccagccc aagctacttg agaatatttt tcatcatttt catagttaca
     4381 ttcttgattg ggttgcttaa agtcctgatc attagacagg tgatactaca atggaatagt
     4441 aataaaatta agtcctcatc agattacaga gtgtcagcct caaaaaagga taagttgatt
     4501 ctgcaaagtg tttgcacaag agcagtcacc taccgacgtg agaagcctga agaaataaaa
     4561 atggatatca gcaaattaaa tgctcatgaa actttcaggt gcaacttcgg atccgactac
     4621 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     4681 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4741 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     4801 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4861 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4921 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     4981 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     5041 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     5101 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     5161 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     5221 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     5281 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     5341 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5401 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5461 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5521 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5581 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5641 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5701 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5761 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5821 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5881 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5941 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6001 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6061 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6121 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6181 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6241 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6301 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6361 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6421 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6481 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6541 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6601 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6661 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6721 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6781 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6841 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6901 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6961 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7021 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     7081 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     7141 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7201 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7261 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7321 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7381 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7441 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7501 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7561 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7621 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7681 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7741 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7801 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7861 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7921 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7981 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8041 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8101 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8161 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8221 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8281 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8341 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8401 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8461 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8521 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8581 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8641 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8701 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8761 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8821 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8881 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8941 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9001 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9061 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9121 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9181 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9241 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     9301 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     9361 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     9421 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9481 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9541 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9601 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9661 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9721 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
