LOCUS       V037871                 9002 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037871
VERSION     V037871
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      2288..2297
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2297..2362
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2369..3904
                     /label="Slc16a14(NM_027921)"
                     /note="Slc16a14(NM_027921)"
                     /gene="Slc16a14"
     promoter        3972..4183
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4196..4464
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             4491..5090
                     /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    5097..5685
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5759..5992
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6064..6185
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6204..6339
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6377..6393)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6401..6417
                     /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(6425..6455)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6470..6491
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6779..7367)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7538..8398)
                     /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(8399..8503)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8977..8993
                     /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 agagctagcc accatggact acaaagacca tgacggtgat tataaagatc atgacatcga
     2341 ctacaaggat gacgatgaca aggaattcat gtatacgagt catgaagata tcgggtatga
     2401 ccttgaagac gaccgaaagg ctaagaataa gaagacgctg aagccccacc cagatatcga
     2461 tggcgggtgg gcctggatga tggtcctctc gtccttcttt gttcacatcc tcatcatggg
     2521 ctcacagatg gccctaggag tcctcaacgt ggagtggctt gaagagtttc accagagccg
     2581 cggcctgacc gcctgggtca gctccctgag catgggcatc accttgatcg tgggaccttt
     2641 catcggcttg ttcattaaca cctgcgggtg ccgccagaca gcaatcattg gaggcctggt
     2701 gaactccctc ggctgggtgt tgagcgccta tgcagccaat gtgcagtctc tctttattac
     2761 ctttggagtg gcagcaggcc ttggcagtgg gatggcttac ctacctgcag tggtcatggt
     2821 gggaaggtac tttcagaaga gacgagccct ggcccagggc ctcagtacca cggggacagg
     2881 atttgggacg ttcctgatga ccgtgttgct gaaatacctg tgcgcagaat atggctggcg
     2941 gaatgctatg ttcatccaag gtgctctgtc cctgaacctg tgtgtctgtg gggcgctcat
     3001 gaggcccctc tctcccgaga agttagaaaa ctgcccagaa gcggaagagc cgtgtgctct
     3061 cccagcttac tccactgaat ctgttaagtc tggaggacca ctgggcatgg ctgaagaaca
     3121 ggacagaagg cccgggaatg aggagatggt gtgtgacctt caaacacagg agtgtcaagg
     3181 tcaaacccat ccgaggaaga atgtgtgtgc cttccgggtt ctgaagacag tgagccagct
     3241 cactgtgcaa gtccggaggg gctttaggga ctggcactca ggctattttg ggacggcttc
     3301 actcttcacc aaccgcatgt ttgtagcctt tattttctgg gctctgttcg catacagcag
     3361 ctttgtcatc cctttcatcc acctgcccga aatcgtcagt ttgtataact tgtcagagca
     3421 aaatgacaca ttccctctga cctcgattat agcaatactt cacatcttcg ggaaggtgat
     3481 cttgggggcc gtggccgacc tcccctgtat cagcgtctgg aatgtcttcc tcatcgctaa
     3541 cttcaccctc gtcctcagca tcttccttct gcctctgatg cacacctacg cgagtctggc
     3601 tgtcatttgt gccctaattg ggttttctag cgggtatttc tccctcatgc ctgtggtgac
     3661 agaggacctg gttggcactg agcacttggc caatgcctac ggcatcatca tctgcgccaa
     3721 tggcatctca gccttactag gaccaccatt cgcaggatgg atctttgaca tcacccaaaa
     3781 atatgacttt tccttttata tatgtggttt gctttacatg gtagggatac tctttttgct
     3841 cattcaaccg tgtattcaga tgatagatca atccagaagg aagtgcatag agggtgcaca
     3901 tgtttaggga tccctcgaga gatcagtgta gcggccgcga aggatctgcg atcgctccgg
     3961 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     4021 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     4081 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     4141 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     4201 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
     4261 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
     4321 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
     4381 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
     4441 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag atgaccgagt
     4501 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg
     4561 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg
     4621 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4681 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4741 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4801 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4861 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4921 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4981 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5041 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga gtcgacaatc
     5101 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     5161 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5221 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5281 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5341 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5401 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5461 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5521 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5581 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5641 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga
     5701 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     5761 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc
     5821 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5881 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5941 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     6001 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     6061 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6121 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6181 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc
     6241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6301 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc
     6361 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6421 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6481 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6541 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6601 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6661 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6721 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6781 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6841 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6901 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6961 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7021 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7081 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7141 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7201 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7261 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7321 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7381 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7441 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7501 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7561 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7621 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7681 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7741 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7801 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7861 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7921 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7981 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     8041 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     8101 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     8161 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8221 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8281 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8341 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8401 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8461 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8521 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     8581 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     8641 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     8701 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     8761 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     8821 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     8881 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     8941 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     9001 tg
//
