LOCUS       V036822                 9136 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V036822
VERSION     V036822
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..4047
                     /label="TBC1D3(NM_001123391)"
                     /note="TBC1D3(NM_001123391)"
                     /gene="TBC1D3"
     promoter        4106..4317
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4330..4598
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4625..5224
                     /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    5231..5819
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5893..6126
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6198..6319
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6338..6473
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6511..6527)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6535..6551
                     /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(6559..6589)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6604..6625
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6913..7501)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7672..8532)
                     /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(8533..8637)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9111..9127
                     /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 atggacgtgg tagaggtcgc gggcagttgg tgggcacaag agcgagagga catcattatg
     2461 aaatacgaaa agggacaccg agctgggctg ccagaggaca aggggcctaa gccttttcga
     2521 agctacaaca acaacgtcga tcatttgggg attgtacatg agacggagct gcctcctctg
     2581 actgcgcggg aggcgaagca aattcggcgg gagatcagcc gaaagagcaa gtgggtggat
     2641 atgctgggag actgggagaa atacaaaagc agcagaaagc tcatagatcg agcgtacaag
     2701 ggaatgccca tgaacatccg gggcccgatg tggtcagtcc tcctgaacac tgaggaaatg
     2761 aagttgaaaa accccggaag ataccagatc atgaaggaga agggcaagag gtcatctgag
     2821 cacatccagc gcatcgaccg ggacgtaagc gggacattaa ggaagcatat attcttcagg
     2881 gatcgatacg gaaccaagca gcgggaacta ctccacatcc tcctggcata tgaggagtac
     2941 aacccggagg tgggctactg cagggacctg agccacatcg ccgccttgtt cctcctctat
     3001 cttcctgagg aggatgcatt ctgggcactg gtgcagctgc tggccagtga gaggcactcc
     3061 ctgcagggat ttcacagccc aaatggcggg accgtccagg ggctccaaga ccaacaggag
     3121 catgtggtag ccacgtcaca acccaagacc atggggcatc aggacaagaa agatctatgt
     3181 gggcagtgtt ccccgttagg ctgcctcatc cggatattga ttgacgggat ctctctcggg
     3241 ctcaccctgc gcctgtggga cgtgtatctg gtagaaggcg aacaggcgtt gatgccgata
     3301 acaagaatcg cctttaaggt tcagcagaag cgcctcacga agacgtccag gtgtggcccg
     3361 tgggcacgtt tttgcaaccg gttcgttgat acctgggcca gggatgagga cactgtgctc
     3421 aagcatctta gggcctctat gaagaaacta acaagaaaga agggggacct gccaccccca
     3481 gccaaacccg agcaagggtc gtcggcatcc aggcctgtgc cggcttcacg tggcgggaag
     3541 accctctgca agggggacag gcaggcccct ccaggcccac cagcccggtt cccgcggccc
     3601 atttggtcag cttccccgcc acgggcacct cgttcttcca caccctgtcc tggtggggct
     3661 gtccgggaag acacctaccc tgtgggcact cagggtgtgc ccagcccggc cctggctcag
     3721 ggaggacctc agggttcctg gagattcctg cagtggaact ccatgccccg cctcccaacg
     3781 gacctggacg tagagggccc ttggttccgc cattatgatt tcaggcagag ctgctgggtc
     3841 cgtgccatat cccaggagga ccagctggcc ccctgctggc aggctgaaca ccctgcggag
     3901 cgggtgagat cggctttcgc tgcacccagc actgattccg accagggcac ccccttcaga
     3961 gctagggacg aacagcagtg tgctcccacc tcagggcctt gcctctgcgg cctccacttg
     4021 gaaagttctc agttccctcc aggcttctag ggatccgata tctagcggcc gcgaaggatc
     4081 tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4141 ttggggggag gggtcggcaa ttgaacgggt gcctagagaa ggtggcgcgg ggtaaactgg
     4201 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4261 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacagctg
     4321 aagcttcgag gggctcgcat ctctccttca cgcgcccgcc gccctacctg aggccgccat
     4381 ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg gtgcctcctg aactgcgtcc
     4441 gccgtctagg taagtttaaa gctcaggtcg agaccgggcc tttgtccggc gctcccttgg
     4501 agcctaccta gactcagccg gctctccacg ctttgcctga ccctgcttgc tcaactctac
     4561 gtctttgttt cgttttctgt tctgcgccgt tacagatcca agctgtgacc ggcgcctacg
     4621 ctagatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
     4681 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
     4741 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4801 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4861 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4921 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     4981 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5041 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5101 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5161 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5221 ctgagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     5281 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     5341 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     5401 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     5461 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     5521 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     5581 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc
     5641 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     5701 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     5761 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     5821 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5881 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     5941 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     6001 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     6061 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     6121 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     6181 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6241 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     6301 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     6361 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6421 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     6481 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6541 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6601 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6661 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6721 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     7021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     7081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     7141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     7201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     7261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     7321 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     7381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     7441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     7501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     7561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     7621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     7681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     7741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     8041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     8101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     8161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     8221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     8281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     8341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     8401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     8461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     8521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     8581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     8641 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     8701 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     8761 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     8821 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     8881 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     8941 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     9001 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     9061 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     9121 ggccagtgcc aagctg
//
