LOCUS       V036927                 9934 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036927
VERSION     V036927
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..4845
                     /label="RABGAP1L(NM_014857)"
                     /note="RABGAP1L(NM_014857)"
                     /gene="RABGAP1L"
     promoter        4904..5115
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5128..5396
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5423..6022
                     /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    6029..6617
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6691..6924
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6996..7117
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7136..7271
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7309..7325)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7333..7349
                     /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(7357..7387)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7402..7423
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7711..8299)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8470..9330)
                     /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(9331..9435)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9909..9925
                     /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 atggaggtca gagcttcatt acagaaggtt agtggatcat ctgattctgt ggctacaatg
     2461 aacagtgaag aatttgtttt ggttcctcag tatgcagatg ataattctac aaaacatgaa
     2521 gaaaaacctc aactgaagat agtttctaat ggtgatgaac aattggaaaa agccatggaa
     2581 gagattttga gagattccga gaaaaggcca agcagtcttc ttgttgattg tcaaagttcc
     2641 agtgagattt cagaccattc gtttggagat attccagcca gccaaacaaa taagccatct
     2701 cttcagttaa ttttggatcc gtctaacaca gaaatttcta cacccagacc atcttctcca
     2761 ggtggactac ctgaagaaga tagtgtttta tttaataaac tgacctactt aggatgtatg
     2821 aaggtttctt ccccacgtaa tgaagtagag gctttacggg caatggcaac catgaaatct
     2881 tccagtcaat acccctttcc tgttaccctg tatgtaccaa atgttccaga aggttctgtg
     2941 agaattatag accaatccag caatgtggag atagcatctt ttccaatcta taaggtgtta
     3001 ttctgtgcac gtggacatga cggaacaaca gagagcaatt gctttgcatt tacagagagt
     3061 tcccatggtt cggaagaatt tcagatacat gttttctcct gtgaaattaa agaggcagta
     3121 agcagaattt tgtacagttt ctgtacagca ttcaaacgtt cttccagaca agtgtctgat
     3181 gttaaagact cagttattcc tacccccgac agtgatgtgt ttaccttcag tgtctccttg
     3241 gaggtaaaag aagacgatgg aaaaggaaac tttagccctg tgcctaagga tagagataaa
     3301 ttttatttca aattaaagca aggaatagag aagaaggttg tgattacagt gcagcaactt
     3361 tctaacaaag aattagctat tgaaagatgt tttggaatgt tattaagccc aggtcgaaac
     3421 gtgaagaaca gtgacatgca tttactggat atggaatcca tgggaaagag ctatgatggg
     3481 agagcttatg tcatcactgg catgtggaac cccaatgcac cagtatttct ggcacttaac
     3541 gaggaaaccc caaaagataa gcaagtatac atgactgtgg cagtggatat ggtagtcaca
     3601 gaggtggtgg agcctgttcg ctttctcctg gagacagtag tccgtgtgta ccctgcaaat
     3661 gagcgatttt ggtatttcag cagaaagact ttcacagaga ctttcttcat gagattgaaa
     3721 cagtctgagg gaaaaggcca taccaatgct ggagatgcaa tatatgaggt ggtgagtcta
     3781 cagcgagagt ctgacaagga ggaaccagtc actcctacta gtggaggggg tccaatgtca
     3841 ccccaggatg atgaagcaga agaggagagt gataatgaac tctcaagtgg aacaggtgat
     3901 gtgtctaagg attgtcctga gaagatcctg tattcttggg gagagttgct aggaaaatgg
     3961 cacagtaacc ttggtgcacg accgaaaggg ctgtctactc tggtgaagag tggtgtccct
     4021 gaagcattga gggcagaggt atggcagtta ttggcaggct gccatgacaa ccaggcaatg
     4081 ctggatagat accgaattct tatcacaaag gactcagccc aggagagtgt tattactcga
     4141 gatattcatc gtacatttcc cgcacatgat tactttaaag atactggagg agatggtcaa
     4201 gaatcgctct ataagatctg caaggcctac tctgtgtatg atgaagacat tgggtactgt
     4261 caagggcagt cttttcttgc tgctgtatta ctgctgcata tgccagagga acaagcattc
     4321 tgtgttttgg tgaaaatcat gtacgactat ggtttgagag acctctacag aaacaacttc
     4381 gaagatcttc attgcaaatt ctaccagttg gagagactaa tgcaggaaca gctaccggac
     4441 ctgcatagcc atttttctga tctgaacctg gaagctcata tgtatgcatc ccagtggttt
     4501 ctcactcttt ttactgccaa gttcccactc tgcatggtgt tccacatcat tgacttactg
     4561 ctttgtgagg gtttgaacat aatctttcat gtagctttgg ctctcctaaa gacctcaaag
     4621 gaagaccttc tgcaggctga ttttgaaggt gctttaaagt tctttagagt tcagcttcca
     4681 aagagataca gggcagagga aaatgcaaga agactgatgg agcaggcttg caatattaaa
     4741 gtaccaacca agaagctgaa gaaatatgag aaagaatatc agacaatgcg agagagtcag
     4801 ctgcaacagg aagacccaat ggatagatac aagtttgtat atttgtaggg atccgatatc
     4861 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4921 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     4981 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     5041 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     5101 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     5161 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     5221 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     5281 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     5341 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     5401 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     5461 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     5521 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     5581 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     5641 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5701 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5761 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5821 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5881 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5941 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     6001 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     6061 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6121 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6181 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6241 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6301 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6361 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6421 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6481 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6541 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6601 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6661 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6721 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6781 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6841 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6901 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6961 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     7021 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     7081 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     7141 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7201 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     7261 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     7321 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     7381 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     7441 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     7501 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     7561 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     7621 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7681 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7741 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7801 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7861 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7921 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7981 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     8041 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     8101 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     8161 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     8221 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     8281 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     8341 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     8401 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     8461 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     8521 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     8581 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     8641 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8701 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8761 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8821 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8881 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8941 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     9001 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     9061 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     9121 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     9181 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     9241 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     9301 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     9361 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     9421 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     9481 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     9541 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     9601 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9661 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9721 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9781 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9841 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9901 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
