LOCUS       V035670                 9274 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035670
VERSION     V035670
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..4128
                     /label="AFP(NM_001134)"
                     /note="AFP(NM_001134)"
                     /gene="AFP"
     CDS             4135..4200
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4244..4455
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4468..4736
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4763..5362
                     /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    5369..5957
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6031..6264
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6336..6457
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6476..6611
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6649..6665)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6673..6689
                     /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(6697..6727)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6742..6763
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7051..7639)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7810..8670)
                     /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(8671..8775)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9249..9265
                     /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 catgaagtgg gtggaatcaa tttttttaat tttcctacta
     2341 aattttactg aatccagaac actgcataga aatgaatatg gaatagcttc catattggat
     2401 tcttaccaat gtactgcaga gataagttta gctgacctgg ctaccatatt ttttgcccag
     2461 tttgttcaag aagccactta caaggaagta agcaaaatgg tgaaagatgc attgactgca
     2521 attgagaaac ccactggaga tgaacagtct tcagggtgtt tagaaaacca gctacctgcc
     2581 tttctggaag aactttgcca tgagaaagaa attttggaga agtacggaca ttcagactgc
     2641 tgcagccaaa gtgaagaggg aagacataac tgttttcttg cacacaaaaa gcccactcca
     2701 gcatcgatcc cacttttcca agttccagaa cctgtcacaa gctgtgaagc atatgaagaa
     2761 gacagggaga cattcatgaa caaattcatt tatgagatag caagaaggca tcccttcctg
     2821 tatgcaccta caattcttct ttgggctgct cgctatgaca aaataattcc atcttgctgc
     2881 aaagctgaaa atgcagttga atgcttccaa acaaaggcag caacagttac aaaagaatta
     2941 agagaaagca gcttgttaaa tcaacatgca tgtgcagtaa tgaaaaattt tgggacccga
     3001 actttccaag ccataactgt tactaaactg agtcagaagt ttaccaaagt taattttact
     3061 gaaatccaga aactagtcct ggatgtggcc catgtacatg agcactgttg cagaggagat
     3121 gtgctggatt gtctgcagga tggggaaaaa atcatgtcct acatatgttc tcaacaagac
     3181 actctgtcaa acaaaataac agaatgctgc aaactgacca cgctggaacg tggtcaatgt
     3241 ataattcatg cagaaaatga tgaaaaacct gaaggtctat ctccaaatct aaacaggttt
     3301 ttaggagata gagattttaa ccaattttct tcaggggaaa aaaatatctt cttggcaagt
     3361 tttgttcatg aatattcaag aagacatcct cagcttgctg tctcagtaat tctaagagtt
     3421 gctaaaggat accaggagtt attggagaag tgtttccaga ctgaaaaccc tcttgaatgc
     3481 caagataaag gagaagaaga attacagaaa tacatccagg agagccaagc attggcaaag
     3541 cgaagctgcg gcctcttcca gaaactagga gaatattact tacaaaatgc gtttctcgtt
     3601 gcttacacaa agaaagcccc ccagctgacc tcgtcggagc tgatggccat caccagaaaa
     3661 atggcagcca cagcagccac ttgttgccaa ctcagtgagg acaaactatt ggcctgtggc
     3721 gagggagcgg ctgacattat tatcggacac ttatgtatca gacatgaaat gactccagta
     3781 aaccctggtg ttggccagtg ctgcacttct tcatatgcca acaggaggcc atgcttcagc
     3841 agcttggtgg tggatgaaac atatgtccct cctgcattct ctgatgacaa gttcattttc
     3901 cataaggatc tgtgccaagc tcagggtgta gcgctgcaaa caatgaagca agagtttctc
     3961 attaaccttg tgaagcaaaa gccacaaata acagaggaac aacttgaggc tgtcattgca
     4021 gatttctcag gcctgttgga gaaatgctgc caaggccagg aacaggaagt ctgctttgct
     4081 gaagagggac aaaaactgat ttcaaaaact cgtgctgctt tgggagttgg atccgactac
     4141 aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag
     4201 tagcggccgc gaaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg
     4261 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg
     4321 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     4381 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     4441 gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc
     4501 cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt
     4561 gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt
     4621 tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc
     4681 ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag
     4741 ctgtgaccgg cgcctacgct agatgaccga gtacaagccc acggtgcgcc tcgccacccg
     4801 cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     4861 gcgccacacc gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     4921 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     4981 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     5041 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     5101 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     5161 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     5221 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     5281 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     5341 gacccgcaag cccggtgcct gagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5401 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5461 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     5521 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     5581 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     5641 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     5701 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     5761 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     5821 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     5881 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     5941 ttgggccgcc tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     6001 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaaaat
     6061 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     6121 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     6181 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     6241 agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat
     6301 aacttgcaaa gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg
     6361 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6421 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc
     6481 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     6541 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     6601 ttttggaggc ctagactttt gcagagacgg cccaaattcg taatcatggt catagctgtt
     6661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6721 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6781 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6841 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6901 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6961 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     8821 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     8881 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     8941 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     9001 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     9061 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     9121 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     9181 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     9241 acgacgttgt aaaacgacgg ccagtgccaa gctg
//
