LOCUS       V031729                 9034 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031729
VERSION     V031729
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..3888
                     /label="Rai2(NM_198409)"
                     /note="Rai2(NM_198409)"
                     /gene="Rai2"
     CDS             3895..3960
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4004..4215
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4228..4496
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4523..5122
                     /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    5129..5717
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5791..6024
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6096..6217
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6236..6371
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6409..6425)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6433..6449
                     /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(6457..6487)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6502..6523
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6811..7399)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7570..8430)
                     /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(8431..8535)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9009..9025
                     /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 catggatgac ctgcagtccc agaatctttc catggacatg
     2341 actgactccc ctcccacttt ggctaataac agactggaga accgcatggc ccagctgata
     2401 actaccgagg cctggaacat caactccact gacctggtaa agaaggccct ggtgaccgtt
     2461 ccggccccat caattctgaa ccccccagct gagtcccaga gtggcatggc tctgaaggta
     2521 gcagccaccg tgctgcagcc cctgtgcctt ggggaaagcc cagtggtgat gcccattcac
     2581 atgcaggtgg agggaagctc tgcgccagag ctcaacccca atggcaacgc tacctacgtt
     2641 atgacgacac aaggccccgt gcagctacca gtggtgttgg aacagcatgt tttccagcac
     2701 ctcaactccc ctctagtcct gccacaggag gccccgtgct cctccaatgc tatccacaac
     2761 aacctcttcc agggggctga ggactctgag gcccagcctc agctcctgga cctgaggatc
     2821 ccaagtcagc cacaggagcc cgcattgcct tttgaagctg tgctccagaa tttattcccc
     2881 acccaaggct ctctgggccc tccaccctgc cagcctcctc ctggctatgc tccagtgcct
     2941 ccccagccct ttaactcccc cttgtccccc ctagtcccgc cagctaccct cttggtgccc
     3001 tatcctgtga ttgtcccctt gccagttcca gtgcccatcc ccatccctgt cccagtgcct
     3061 cagagtactg aatccaagtt cagccccact ttccccaagc cgccatcttc cttcggcttg
     3121 cactccttta aaggcaccca gaccactctg gaaaaggacg aactgaagcc cttagacatc
     3181 ctccagtcaa aggagtattt ccagctcagc cgccacacag tcatcaagat ggggagtgag
     3241 aatgaggccc tggatctgtc catgaagtca gtgccctggc tcaaggctgg ggaagctagc
     3301 cccccagtct ttcaagagga tgcagcccta gatctgtctt tggcagccca ccgaaaagct
     3361 gaggctcctc cagagacatt gtataacaac agtgggtcag cagacatcca gggtcacacc
     3421 atactggaga aactttccag tggcatggaa atgccctttg cccctgccaa gtgttctgaa
     3481 gcctcagcca tgatggagag ccacagcagc aacagcaacg gcactgagat ggtcagtcag
     3541 cccagccatc ctggcagtga gctgaaggct gaaaataaca ttgagatctt aagtgagtct
     3601 caggcggcca aggtcattgt gtcagttgaa gacgccgtgc ctgccatctt ctgtggcaag
     3661 attaaaggcc tctcaggggt atccaccaaa aacttctcct tcaaaagaga agactctgtg
     3721 cttcagggtt atgatatcaa cagccaagga gaagagtccc tgggaaacgc cgagcccctt
     3781 aggaaaccca tcaaaaaccg gagcataaag ttaaagaaag tgaactccca ggaaatacac
     3841 atgcttccaa ttaaaaaaca acggctggcc acctttttcc caagaaaggg atccgactac
     3901 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     3961 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4021 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     4081 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4141 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4201 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     4261 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     4321 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     4381 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     4441 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     4501 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4561 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4621 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4681 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4741 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     4801 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     4861 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     4921 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     4981 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5041 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5101 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5161 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5221 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5281 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5341 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5401 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5461 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5521 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5581 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5641 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5701 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     5761 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     5821 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     5881 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     5941 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6001 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6061 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6121 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6181 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6241 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6301 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6361 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6421 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6481 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6541 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6601 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6661 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6721 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6781 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6841 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6901 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6961 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7021 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7081 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7141 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7201 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7261 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7321 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7381 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7441 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7501 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7561 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7621 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7681 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7741 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7801 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7861 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7921 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7981 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8041 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8101 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8161 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8221 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8281 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8341 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8401 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8461 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8521 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8581 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8641 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8701 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     8761 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     8821 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     8881 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     8941 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9001 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
