LOCUS       V036845                 9433 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036845
VERSION     V036845
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"
     regulatory      2296..2305
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2305..2394
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             2401..4344
                     /label="TBC1D17(NM_024682)"
                     /note="TBC1D17(NM_024682)"
                     /gene="TBC1D17"
     promoter        4403..4614
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4627..4895
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4922..5521
                     /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    5528..6116
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6190..6423
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6495..6616
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6635..6770
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6808..6824)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6832..6848
                     /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(6856..6886)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6901..6922
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7210..7798)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7969..8829)
                     /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(8830..8934)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9408..9424
                     /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 catggaacaa aaactaatat cagaggaaga tttggaacag
     2341 aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga cttactcgag
     2401 atggaaggag ccggctacag ggtggtgttt gagaagggcg gagtgtacct gcacaccagc
     2461 gctaagaagt atcaggaccg agactctctc atcgctggtg tcatccgtgt cgtggaaaag
     2521 gacaatgacg tcctcctgca ctgggctcct gtagaggagg ctggagattc cacccaaatc
     2581 ctcttctcca agaaggactc cagtgggggt gactcatgtg cttctgagga ggaaccaacc
     2641 tttgaccccg gctatgaacc tgactgggct gtcatcagca ctgtgcggcc acagctctgc
     2701 cactcagagc ccacgagagg tgcagagccc agctgccccc agggctcctg ggccttctca
     2761 gtgagtctgg gggagctaaa gtccatccgc cgctccaagc caggcctcag ctgggcctac
     2821 ctggttctgg tgacccaggc tggaggttcc ctgcccgcac tgcacttcca ccgcgggggc
     2881 acccgcgccc tgctccgcgt cctcagccgc tacctgctgt tggccagctc cccgcaggac
     2941 tcccgcctct accttgtctt cccccacgac tcctctgctc tctccaactc cttccaccac
     3001 ctgcagctct ttgaccagga cagctccaat gtggtgtcac gcttcctcca ggatccctac
     3061 tccaccacct tcagcagctt ctcccgagtg accaacttct tccggggtgc cctgcagcca
     3121 cagcctgagg gagccgcctc cgaccttccc ccgccacccg acgatgagcc cgagcctgga
     3181 ttcgaggtca tttcctgtgt ggagctgggg cctcggccaa ccgtggagcg gggccctcca
     3241 gttacagagg aggagtgggc acgccacgtg ggccctgaag gtcgcctgca gcaggtccct
     3301 gagctgaaga accggatctt ctcggggggt ctgagcccca gcctgcggcg cgaggcctgg
     3361 aagttcctcc tagggtacct cagctgggaa ggcacagctg aggagcacaa ggcccacata
     3421 cgcaagaaaa cggatgagta tttccgcatg aagctgcagt ggaaatctgt gagccctgag
     3481 caggagcgga gaaactcact tctgcatgga taccgcagcc tcatcgaaag ggatgtgagc
     3541 cgcactgaca ggaccaacaa gttctacgag ggtcccgaga acccggggct gggcctgctg
     3601 aacgatatcc tcctcaccta ctgcatgtat cacttcgacc tcggctacgt ccagggcatg
     3661 agtgatcttc tctccccgat cctctacgtc attcagaacg aggtggatgc tttctggtgt
     3721 ttctgtggct tcatggagct cgtgcaaggg aactttgaag agagccagga gaccatgaag
     3781 cggcaactcg ggcgactgct gctgctcctg agggtgctgg accccctgct ctgcgacttc
     3841 ctggattccc aggactccgg ctctctctgc ttctgtttcc ggtggctgct catctggttc
     3901 aagagggaat tccccttccc ggatgtcctt cggctgtggg aggtgctgtg gacagggctc
     3961 cctggcccca atctgcacct gctggtggcc tgcgccatcc tggacatgga gagggacacc
     4021 ctcatgctgt ccggcttcgg ctccaatgag atcctcaagc acatcaacga gctgactatg
     4081 aagctgagcg tggaggacgt gctgacccgc gccgaggccc tgcaccgcca gctaaccgcc
     4141 tgccccgagc tgccccacaa cgtgcaggag atcctggggc tggccccgcc cgcagagccc
     4201 cacagcccct cgcccaccgc ctccccgctg cctctgtcgc ccacccgggc cccgcccacc
     4261 ccgccgccct ccacggacac agccccgcag cccgacagca gcctggagat cctgcccgag
     4321 gaggaggacg agggcgccga ctcctaagga tccgatatct agcggccgcg aaggatctgc
     4381 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     4441 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     4501 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     4561 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     4621 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     4681 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     4741 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     4801 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     4861 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     4921 gatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     4981 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     5041 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     5101 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     5161 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     5221 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     5281 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     5341 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     5401 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     5461 cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg
     5521 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5581 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5641 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5701 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5761 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5821 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     5881 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     5941 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6001 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6061 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6121 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     6181 aggggggact ggaagggcta attcactccc aacgaaaata agatctgctt tttgcttgta
     6241 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     6301 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     6361 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     6421 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
     6481 cagagagtga gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6541 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6601 tcatcaatgt atcttatcat gtctggctct agctatcccg cccctaactc cgcccagttc
     6661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagacttttg
     6781 cagagacggc ccaaattcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     7021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     7081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     7141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     7201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     7261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     7321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     7381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     7441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     7501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     7561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     7621 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     7681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     7741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7861 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7921 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7981 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     8041 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     8101 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     8161 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     8221 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     8281 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     8341 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     8401 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     8461 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     8521 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     8581 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     8641 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     8701 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     8761 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8821 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8881 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8941 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     9001 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     9061 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     9121 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     9181 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     9241 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     9301 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     9361 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     9421 cagtgccaag ctg
//
