LOCUS       V032516                 9223 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032516
VERSION     V032516
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..4077
                     /label="Igf2bp2(NM_183029)"
                     /note="Igf2bp2(NM_183029)"
                     /gene="Igf2bp2"
     CDS             4084..4149
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4193..4404
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4417..4685
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4712..5311
                     /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    5318..5906
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5980..6213
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6285..6406
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6425..6560
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6598..6614)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6622..6638
                     /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(6646..6676)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6691..6712
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7000..7588)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7759..8619)
                     /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(8620..8724)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9198..9214
                     /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 catgatgaac aagctgtaca ttgggaacct gagccccgcc
     2341 gtcaccgccg acgacctccg gcagctcttc ggggacagga agctgcccct ggcgggacag
     2401 gtcctactca agtccggcta cgccttcgtg gactaccccg accagaactg ggccatccgc
     2461 gccatcgaga ccctctcggg taaagtggaa ttgcatggga aaatcatgga agttgactac
     2521 tcagtctcta aaaagctaag gagcagaaga atccagattc ggaacatccc gcctcacctg
     2581 cagtgggagg tgttggatgg gctgttggct gaatatggga cagtggagaa cgtggagcaa
     2641 gtcaacacag atacagaaac tgccgttgtc aacgtcacct atatgacaag agaagaggca
     2701 aagctagcta ttgagaagct cagtgggcat cagtttgagg actactcctt caagatttcc
     2761 tacatccccg atgaagaggt gagctctcct tcaccccctc atcgtgcccg ggaacaaggc
     2821 cacggccccg ggagctcttc tcaggccaga cagattgatt tcccgctgcg gatcctggtc
     2881 cccacccagt ttgttggtgc catcatcgga aaggagggct tgaccataaa gaacatcact
     2941 aagcagaccc agtcccgggt agacatccac agaaaggaga actctggggc tgcagagaag
     3001 cctgtcacaa tccatgctac cccagaaggg acatctgaag catgccgcat gattcttgag
     3061 attatgcaaa aagaagctga tgagaccaaa ctggctgagg aggttcctct gaaaatcctg
     3121 gcccacaatg gcttcgttgg aagactgatt ggcaaagaag gcagaaacct gaagaaaata
     3181 gaacatgaga cagggaccaa gataaccatc tcatccttgc aggatttgag catttataac
     3241 cccgagagaa ccatcaccgt gaggggcacc attgaagcct gtgccaatgc tgagatagag
     3301 attatgaaga agctccgaga ggcctttgag aacgacatgc tggccgttaa ccaacaagcc
     3361 aatctgatcc cagggctaaa cctcagtgca cttggcatct tttcgactgg actgtctgtg
     3421 cttcctccac cagcagggcc ccgtggagtt ccccccagtc ctccctatca cccctttgct
     3481 acccactccg gatacttctc cagtctgtac cctcatcacc atttcggccc attcccacat
     3541 catcactcct acccagagca ggagactgta agtctcttca tcccaaccca ggctgtgggt
     3601 gctatcatcg ggaagaaggg ggcacacatc aaacagctcg ctcgatttgc tggtgcctcc
     3661 atcaagattg ctccagcaga aggtccagat gtcagtgaga ggatggtcat catcactggt
     3721 cctcctgaag cccagtttaa ggctcaggga cggatctttg ggaaactgaa ggaagaaaac
     3781 ttctttaatc ccaaagaaga agtgaagctg gaggcccaca tccgagtccc ctcgtcgacc
     3841 gctggccggg tgattggcaa gggcgggaaa accgtgaacg agctgcagaa cttgacaagt
     3901 gcagaagtta tcgtgcctcg tgaccaaacg ccagacgaga atgaggaagt gatcgtcaga
     3961 attatcgggc atttttttgc tagccagact gcacaacgca agatcaggga aattgtacag
     4021 caggtgaagc agcaggagca gagataccct cagggagtcg ccccacagcg cagcaaggga
     4081 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     4141 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     4201 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     4261 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     4321 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     4381 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     4441 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     4501 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     4561 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     4621 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     4681 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     4741 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     4801 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     4861 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     4921 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     4981 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5041 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5101 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5161 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     5221 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     5281 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat
     5341 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     5401 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     5461 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     5521 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     5581 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     5641 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     5701 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     5761 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     5821 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     5881 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     5941 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6001 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6061 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6121 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6181 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     6241 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     6301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     6421 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6481 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6541 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     6601 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     6661 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     6721 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6781 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     6841 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6901 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6961 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7021 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7081 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7141 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7201 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7261 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7321 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7381 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7441 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7501 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7561 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     7621 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     7681 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     7741 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     7801 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     7861 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     7921 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     7981 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8041 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8101 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8161 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8221 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8281 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8341 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8401 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8461 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8521 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     8581 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     8641 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     8701 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     8761 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     8821 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     8881 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     8941 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9001 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9061 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9121 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9181 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
