LOCUS       V023701                 9024 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023701
VERSION     V023701
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             2301..3878
                     /label="FTO(NM_001363894)"
                     /note="FTO(NM_001363894)"
                     /gene="FTO"
     CDS             3885..3950
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3994..4205
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4218..4486
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4513..5112
                     /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    5119..5707
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5781..6014
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6086..6207
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6226..6361
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6399..6415)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6423..6439
                     /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(6447..6477)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6492..6513
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6801..7389)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7560..8420)
                     /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(8421..8525)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8999..9015
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgaagcgca ccccgactgc cgaggaacga gagcgcgaag
     2341 ctaagaaact gaggcttctt gaagagcttg aagacacttg gctcccttat ctgaccccca
     2401 aagatgatga attctatcag cagtggcagc tgaaatatcc taaactaatt ctccgagaag
     2461 ccagcagtgt atctgaggag ctccataaag aggttcaaga agcctttctc acactgcaca
     2521 agcatggctg cttatttcgg gacctggtta ggatccaagg caaagatctg ctcactccgg
     2581 tatctcgcat cctcattggt aatccaggct gcacctacaa gtacctgaac accaggctct
     2641 ttacggtccc ctggccagtg aaagggtcta atataaaaca caccgaggct gaaatagccg
     2701 ctgcttgtga gaccttcctc aagctcaatg actacctgca gatagaaacc atccaggctt
     2761 tggaagaact tgctgccaaa gagaaggcta atgaggatgc tgtgccattg tgtatgtctg
     2821 cagatttccc cagggttggg atgggttcat cctacaacgg acaagatgaa gtggacatta
     2881 agagcagagc agcatacaac gtaactttgc tgaatttcat ggatcctcag aaaatgccat
     2941 acctgaaaga ggaaccttat tttggcatgg ggaaaatggc agtgagctgg catcatgatg
     3001 aaaatctggt ggacaggtca gcggtggcag tgtacagtta tagctgtgaa ggccctgaag
     3061 aggaaagtga ggatgactct catctcgaag gcagggatcc tgatatttgg catgttggtt
     3121 ttaagatctc atgggacata gagacacctg gtttggcgat accccttcac caaggagact
     3181 gctatttcat gcttgatgat ctcaatgcca cccaccaaca ctgtgttttg gccggttcac
     3241 aacctcggtt tagttccacc caccgagtgg cagagtgctc aacaggaacc ttggattata
     3301 ttttacaacg ctgtcagttg gctctgcaga atgtctgtga cgatgtggac aatgatgatg
     3361 tctctttgaa atcctttgag cctgcagttt tgaaacaagg agaagaaatt cataatgagg
     3421 tcgagtttga gtggctgagg cagttttggt ttcaaggcaa tcgatacaga aagtgcactg
     3481 actggtggtg tcaacccatg gctcaactgg aagcactgtg gaagaagatg gagggtgtga
     3541 taagtgtgga gttatacaaa gagctaggac ttatgtctcc caggttgata aggcacaagg
     3601 caacaaatgc tgtgcttcat gaagttaaaa gagaggggct ccccgtggaa caaaggaatg
     3661 aaatcttgac tgccatcctt gcctcgctca ctgcacgcca gaacctgagg agagaatggc
     3721 atgccaggtg ccagtcacga attgcccgaa cattacctgc tgatcagaag ccagaatgtc
     3781 ggccatactg ggaaaaggat gatgcttcga tgcctctgcc gtttgacctc acagacatcg
     3841 tttcagaact cagaggtcag cttctggaag caaaacccgg atccgactac aaagaccatg
     3901 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagcggccgc
     3961 gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
     4021 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg
     4081 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
     4141 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
     4201 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag
     4261 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa
     4321 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc
     4381 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc
     4441 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg
     4501 cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4561 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4621 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4681 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4741 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4801 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4861 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4921 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4981 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5041 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     5101 cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5521 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5701 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5761 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat aagatctgct
     5821 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5881 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5941 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6001 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6061 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     6121 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6181 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     6241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6301 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6361 ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt tcctgtgtga
     6421 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6481 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6541 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6601 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6661 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6721 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7201 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     7261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7621 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7681 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7741 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7801 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7861 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7921 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7981 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8041 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8101 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8161 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8221 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8281 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8341 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8401 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8461 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8521 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     8581 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     8641 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     8701 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     8761 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     8821 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     8881 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     8941 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     9001 aaaacgacgg ccagtgccaa gctg
//
