LOCUS       V025645                 9183 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025645
VERSION     V025645
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..4037
                     /label="IGF2BP3(NM_006547)"
                     /note="IGF2BP3(NM_006547)"
                     /gene="IGF2BP3"
     CDS             4044..4109
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4153..4364
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4377..4645
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4672..5271
                     /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    5278..5866
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5940..6173
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6245..6366
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6385..6520
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6558..6574)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6582..6598
                     /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(6606..6636)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6651..6672
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6960..7548)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7719..8579)
                     /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(8580..8684)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9158..9174
                     /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 atgaacaaac tgtatatcgg aaacctcagc gagaacgccg
     2341 ccccctcgga cctagaaagt atcttcaagg acgccaagat cccggtgtcg ggacccttcc
     2401 tggtgaagac tggctacgcg ttcgtggact gcccggacga gagctgggcc ctcaaggcca
     2461 tcgaggcgct ttcaggtaaa atagaactgc acgggaaacc catagaagtt gagcactcgg
     2521 tcccaaaaag gcaaaggatt cggaaacttc agatacgaaa tatcccgcct catttacagt
     2581 gggaggtgct ggatagttta ctagtccagt atggagtggt ggagagctgt gagcaagtga
     2641 acactgactc ggaaactgca gttgtaaatg taacctattc cagtaaggac caagctagac
     2701 aagcactaga caaactgaat ggatttcagt tagagaattt caccttgaaa gtagcctata
     2761 tccctgatga aatggccgcc cagcaaaacc ccttgcagca gccccgaggt cgccgggggc
     2821 ttgggcagag gggctcctca aggcaggggt ctccaggatc cgtatccaag cagaaaccat
     2881 gtgatttgcc tctgcgcctg ctggttccca cccaatttgt tggagccatc ataggaaaag
     2941 aaggtgccac cattcggaac atcaccaaac agacccagtc taaaatcgat gtccaccgta
     3001 aagaaaatgc gggggctgct gagaagtcga ttactatcct ctctactcct gaaggcacct
     3061 ctgcggcttg taagtctatt ctggagatta tgcataagga agctcaagat ataaaattca
     3121 cagaagagat ccccttgaag attttagctc ataataactt tgttggacgt cttattggta
     3181 aagaaggaag aaatcttaaa aaaattgagc aagacacaga cactaaaatc acgatatctc
     3241 cattgcagga attgacgctg tataatccag aacgcactat tacagttaaa ggcaatgttg
     3301 agacatgtgc caaagctgag gaggagatca tgaagaaaat cagggagtct tatgaaaatg
     3361 atattgcttc tatgaatctt caagcacatt taattcctgg attaaatctg aacgccttgg
     3421 gtctgttccc acccacttca gggatgccac ctcccacctc agggccccct tcagccatga
     3481 ctcctcccta cccgcagttt gagcaatcag aaacggagac tgttcatctg tttatcccag
     3541 ctctatcagt cggtgccatc atcggcaagc agggccagca catcaagcag ctttctcgct
     3601 ttgctggagc ttcaattaag attgctccag cggaagcacc agatgctaaa gtgaggatgg
     3661 tgattatcac tggaccacca gaggctcagt tcaaggctca gggaagaatt tatggaaaaa
     3721 ttaaagaaga aaactttgtt agtcctaaag aagaggtgaa acttgaagct catatcagag
     3781 tgccatcctt tgctgctggc agagttattg gaaaaggagg caaaacggtg aatgaacttc
     3841 agaatttgtc aagtgcagaa gttgttgtcc ctcgtgacca gacacctgat gagaatgacc
     3901 aagtggttgt caaaataact ggtcacttct atgcttgcca ggttgcccag agaaaaattc
     3961 aggaaattct gactcaggta aagcagcacc aacaacagaa ggctctgcaa agtggaccac
     4021 ctcagtcaag acggaaggga tccgactaca aagaccatga cggtgattat aaagatcatg
     4081 acatcgacta caaggatgac gatgacaagt agcggccgcg aaggatctgc gatcgctccg
     4141 gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg
     4201 tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg
     4261 tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg
     4321 ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg
     4381 ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga
     4441 gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa
     4501 gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac
     4561 tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt
     4621 tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta gatgaccgag
     4681 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc
     4741 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc
     4801 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4861 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4921 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4981 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     5041 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     5101 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     5161 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     5221 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat
     5281 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5341 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5401 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5461 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5521 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5581 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5641 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     5701 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     5761 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     5821 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag
     5881 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     5941 ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta ctgggtctct
     6001 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6061 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6121 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6181 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6241 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6361 atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc cgcccattct
     6421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     6481 gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg cagagacggc
     6541 ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6601 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6661 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6721 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6781 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6841 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6901 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6961 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     7021 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     7081 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7141 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7201 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7261 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7321 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7381 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7441 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7501 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7561 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7621 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7681 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7741 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7801 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7861 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7921 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7981 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     8041 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     8101 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8161 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8221 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8281 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8341 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8401 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8461 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8521 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8581 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8641 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8701 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     8761 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     8821 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     8881 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     8941 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     9001 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     9061 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     9121 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag
     9181 ctg
//
