LOCUS       V039792                10003 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039792
VERSION     V039792
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..4857
                     /label="KIF18B(NM_001265577)"
                     /note="KIF18B(NM_001265577)"
                     /gene="KIF18B"
     CDS             4864..4929
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4973..5184
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5197..5465
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5492..6091
                     /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    6098..6686
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6760..6993
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7065..7186
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7205..7340
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7378..7394)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7402..7418
                     /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(7426..7456)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7471..7492
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7780..8368)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8539..9399)
                     /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(9400..9504)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9978..9994
                     /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 catggcagtg gaggacagca cgctgcaagt agtggtacgg
     2341 gtgcggcccc ccacccctcg ggagctggac agtcagcggc ggccagtggt tcaggtggtg
     2401 gacgagcggg tgctggtgtt taaccctgag gagcccgatg gagggttccc tggcctgaaa
     2461 tggggtggca cccatgatgg ccccaagaag aagggcaaag acctgacgtt tgtctttgac
     2521 cgggtctttg gcgaggcggc cacccaacag gacgtgttcc agcacaccac gcacagcgtc
     2581 ctggacagct tcctccaggg ctacaactgc tcagtgtttg cctacggggc caccggggct
     2641 gggaagacac acaccatgct gggaagggag ggggaccccg gcatcatgta cctgaccacc
     2701 gtggaactgt acaggcgcct ggaggcccgc cagcaggaga agcacttcga ggtgctcatc
     2761 agctaccagg aggtgtataa tgaacagatc catgacctcc tggagcccaa ggggcccctt
     2821 gccatccgcg aggaccccga caagggggtg gtggtgcaag gactttcttt ccaccagcca
     2881 gcctcagccg agcagctgct ggagatactg accaggggga accgtaaccg cacgcagcac
     2941 cccactgatg ccaacgcgac ttcctcccgc tcccatgcca tcttccagat ctttgtgaag
     3001 cagcaggacc gggttccagg actgacccag gctgtccagg tggccaagat gagcctgatt
     3061 gacctggctg gctcagagcg ggcatccagc acccatgcga agggggagcg gctgcgggag
     3121 ggggccaaca tcaaccgctc tctgctggcg ctcatcaacg tcctcaatgc cttggccgat
     3181 gcaaagggcc gcaagaccca tgtgccctac cgggacagca aactgacccg cctgctcaaa
     3241 gactccctcg ggggcaactg ccgcacagtg atgatcgctg ccatcagccc ctccagcctg
     3301 acctacgagg acacgtacaa caccctcaaa tatgccgacc gggccaagga gatcaggctc
     3361 tcgctgaaga gcaatgtgac cagcctggac tgtcacatca gccagtatgc taccatctgc
     3421 caacagctcc aggctgaggt agccgctctg aggaagaagc tccaagtgta tgagggggga
     3481 ggccagcccc caccacagga cctcccagga tctcccaagt cgggaccacc accagaacac
     3541 cagccctgca ccccagagct ccctgcaggg cctagagccc ttcaagagga gagtctgggg
     3601 atggaggccc aggtggagag ggccatggaa gggaactctt cagaccagga gcagtcccca
     3661 gaggatgagg atgaaggccc agctgaggag gttccaaccc agatgccaga gcagaacccc
     3721 acacatgcac tgccagagtc ccctcgcctg accctgcagc ccaagccagt cgtgggccac
     3781 ttctcagcac gggaactgga tggggaccgt tctaagcagt tggccctaaa ggtgctgtgc
     3841 gttgcccagc ggcagtactc cctgctccaa gcagccaacc tcctgacgcc cgacatgatc
     3901 acagagtttg agaccctaca gcagctggtg caagaggaaa aaattgagcc tggggcagag
     3961 gccttgagga cttcaggcct ggccaggggg gcacctctgg ctcaggagct gtgttcagag
     4021 tcaatccctg tgccgtctcc tctctgccca gagcctccag gatacactgg ccctgtgacc
     4081 cggactatgg cgaggcgact gagtggcccc ctgcacaccc tgggaatccc gcctggaccc
     4141 aactgcaccc cagcccaggg gtcccgatgg cccatggaga agaagaggag gagaccaagc
     4201 gccttggagg cagacagtcc catggcccca aagcggggca ccaagcgcca gcgccagtcc
     4261 ttcctgccct gcctaaggag agggtctctg cctgacaccc aaccttcaca ggggcccagc
     4321 acccccaaag gagaaagggc ctcctccccc tgccattccc ctcgcgtttg cccagccaca
     4381 gtcatcaaaa gccgggtgcc cctgggccct tccgccatgc agaactgctc caccccgctg
     4441 gctctgccca ctcgagacct caatgccacc tttgatctct ctgaggagcc tccctcaaag
     4501 cccagtttcc atgaatgcat tggctgggac aaaatacccc aggagctgag caggctggac
     4561 cagcccttca tccccagggc acctgtgccc ctgttcacca tgaagggccc caagccaaca
     4621 tcttccctcc ctgggacctc tgcctgcaag aagaagcgcg ttgcgagttc ctcagtctcc
     4681 catggccgca gccgcatcgc ccgcctcccc agcagcactt tgaagaggcc agctgggccc
     4741 cttgtactcc cagagctgcc cttgagtccc ctgtgcccta gcaaccggag gaatggaaag
     4801 gacctcatca gggtggggag agcactctca gcagggaacg gcgtcaccaa ggtgtccgga
     4861 tccgactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
     4921 gatgacaagt agcggccgcg aaggatctgc gatcgctccg gtgcccgtca gtgggcagag
     4981 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc
     5041 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     5101 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     5161 aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc
     5221 gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc
     5281 gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga
     5341 ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt
     5401 tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac
     5461 agatccaagc tgtgaccggc gcctacgcta gatgaccgag tacaagccca cggtgcgcct
     5521 cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta
     5581 ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca
     5641 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     5701 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     5761 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     5821 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     5881 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     5941 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     6001 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     6061 ctggtgcatg acccgcaagc ccggtgcctg agtcgacaat caacctctgg attacaaaat
     6121 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6181 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6241 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6301 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6361 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6421 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6481 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6541 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6601 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6661 gatctccctt tgggccgcct ccccgcctgg tacctttaag accaatgact tacaaggcag
     6721 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc
     6781 aacgaaaata agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag
     6841 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6901 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6961 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
     7021 agtatttata acttgcaaag aaatgaatat cagagagtga gaggaacttg tttattgcag
     7081 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     7141 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggctct
     7201 agctatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     7261 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     7321 aggaggcttt tttggaggcc tagacttttg cagagacggc ccaaattcgt aatcatggtc
     7381 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7441 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7501 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7561 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7621 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7681 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7741 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7801 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7861 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7921 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7981 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     8041 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     8101 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     8161 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     8221 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     8281 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     8341 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8401 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8461 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8521 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8581 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8641 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8701 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8761 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8821 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8881 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8941 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     9001 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     9061 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     9121 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     9181 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     9241 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     9301 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9361 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9421 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9481 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     9541 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     9601 cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac
     9661 agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt
     9721 tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     9781 ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca
     9841 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
     9901 acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt
     9961 ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctg
//
