LOCUS       V032319                 8234 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032319
VERSION     V032319
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             2287..2352
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2377..3159
                     /label="HSD17B8(NM_014234)"
                     /note="HSD17B8(NM_014234)"
                     /gene="HSD17B8"
     promoter        3204..3415
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3428..3696
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3723..4322
                     /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    4329..4917
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             4991..5224
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5296..5417
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5436..5571
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5609..5625)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5633..5649
                     /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(5657..5687)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5702..5723
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6011..6599)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6770..7630)
                     /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(7631..7735)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8209..8225
                     /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 agaatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2341 gacgatgaca agtctagaga attcggatcc gccaccatgg cgtctcagct ccagaaccga
     2401 ctccgctccg cactggcctt ggtcacaggt gcggggagcg gcatcggccg agcggtcagt
     2461 gtacgcctgg ccggagaggg ggccaccgta gctgcctgcg acctggaccg ggcagcggca
     2521 caggagacgg tgcggctgct gggcgggcca gggagcaagg aggggccgcc ccgagggaac
     2581 catgctgcct tccaggctga cgtgtctgag gccagggccg ccaggtgcct gctggaacaa
     2641 gtgcaggcct gcttttctcg cccaccatct gtcgttgtgt cctgtgcggg catcacccag
     2701 gatgagtttc tgctgcacat gtctgaggat gactgggaca aagtcatagc tgtcaacctc
     2761 aagggcacct tcctagtcac tcaggctgca gcacaagccc tggtgtccaa tggttgtcgt
     2821 ggttccatca tcaacatcag tagcatcgta ggaaaggtgg ggaacgtggg gcagacaaac
     2881 tatgcagcat ccaaggctgg agtgattggg ctgacccaga ccgcagcccg ggagcttgga
     2941 cgacatggga tccgctgtaa ctctgtcctc ccagggttca ttgcaacacc catgacacag
     3001 aaagtgccac agaaagtggt ggacaagatt actgaaatga tcccgatggg acacttgggg
     3061 gaccctgagg atgtggcaga tgtggtcgca ttcttggcat ctgaagatag tggatacatc
     3121 acagggacct cagtggaagt cactggaggt cttttcatgt aagcggccgc gaaggatctg
     3181 cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     3241 ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg taaactggga
     3301 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     3361 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacagctgaa
     3421 gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag gccgccatcc
     3481 acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc
     3541 cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc tcccttggag
     3601 cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc aactctacgt
     3661 ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg cgcctacgct
     3721 agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg
     3781 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg
     3841 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     3901 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     3961 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     4021 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     4081 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     4141 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     4201 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     4261 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     4321 gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     4381 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     4441 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     4501 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     4561 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     4621 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4681 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     4741 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     4801 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     4861 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg
     4921 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     4981 aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct ttttgcttgt
     5041 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5101 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5161 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5221 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
     5281 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5341 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5401 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt
     5461 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5521 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctagactttt
     5581 gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5641 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5701 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6421 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6841 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6961 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7201 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7261 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7441 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7621 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7741 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7801 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     7861 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     7921 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     7981 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     8041 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     8101 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     8161 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     8221 ccagtgccaa gctg
//
