LOCUS       V025533                10020 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025533
VERSION     V025533
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             2301..4874
                     /label="Tas1r3(NM_031872)"
                     /note="Tas1r3(NM_031872)"
                     /gene="Tas1r3"
     CDS             4881..4946
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4990..5201
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             5214..5482
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             5509..6108
                     /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    6115..6703
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6777..7010
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    7082..7203
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      7222..7357
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(7395..7411)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7419..7435
                     /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(7443..7473)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7488..7509
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7797..8385)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8556..9416)
                     /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(9417..9521)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9995..10011
                     /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 tgaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagattcta
     2281 gagctagcga attcgccacc atgccagctt tggctatcat gggtctcagc ctggctgctt
     2341 tcctggagct tgggatgggg gcctctttgt gtctgtcaca gcaattcaag gcacaagggg
     2401 actacatact gggcgggcta tttcccctgg gctcaaccga ggaggccact ctcaaccaga
     2461 gaacacaacc caacagcatc ccgtgcaaca ggttctcacc ccttggtttg ttcctggcca
     2521 tggctatgaa gatggctgtg gaggagatca acaatggatc tgccttgctc cctgggctgc
     2581 ggctgggcta tgacctattt gacacatgct ccgagccagt ggtcaccatg aaatccagtc
     2641 tcatgttcct ggccaaggtg ggcagtcaaa gcattgctgc ctactgcaac tacacacagt
     2701 accaaccccg tgtgctggct gtcatcggcc cccactcatc agagcttgcc ctcattacag
     2761 gcaagttctt cagcttcttc ctcatgccac aggtcagcta tagtgccagc atggatcggc
     2821 taagtgaccg ggaaacgttt ccatccttct tccgcacagt gcccagtgac cgggtgcagc
     2881 tgcaggcagt tgtgactctg ttgcagaact tcagctggaa ctgggtggcc gccttaggga
     2941 gtgatgatga ctatggccgg gaaggtctga gcatcttttc tagtctggcc aatgcacgag
     3001 gtatctgcat cgcacatgag ggcctggtgc cacaacatga cactagtggc caacagttgg
     3061 gcaaggtgct ggatgtacta cgccaagtga accaaagtaa agtacaagtg gtggtgctgt
     3121 ttgcctctgc ccgtgctgtc tactcccttt ttagttacag catccatcat ggcctctcac
     3181 ccaaggtatg ggtggccagt gagtcttggc tgacatctga cctggtcatg acacttccca
     3241 atattgcccg tgtgggcact gtgcttgggt ttttgcagcg gggtgcccta ctgcctgaat
     3301 tttcccatta tgtggagact caccttgccc tggccgctga cccagcattc tgtgcctcac
     3361 tgaatgcgga gttggatctg gaggaacatg tgatggggca acgctgtcca cggtgtgacg
     3421 acatcatgct gcagaaccta tcatctgggc tgttgcagaa cctatcagct gggcaattgc
     3481 accaccaaat atttgcaacc tatgcagctg tgtacagtgt ggctcaagcc cttcacaaca
     3541 ccctacagtg caatgtctca cattgccacg tatcagaaca tgttctaccc tggcagctcc
     3601 tggagaacat gtacaatatg agtttccatg ctcgagactt gacactacag tttgatgctg
     3661 aagggaatgt agacatggaa tatgacctga agatgtgggt gtggcagagc cctacacctg
     3721 tattacatac tgtgggcacc ttcaacggca cccttcagct gcagcagtct aaaatgtact
     3781 ggccaggcaa ccaggtgcca gtctcccagt gttcccgcca gtgcaaagat ggccaggttc
     3841 gccgagtaaa gggctttcat tcctgctgct atgactgcgt ggactgcaag gcgggcagct
     3901 accggaagca tccagatgac ttcacctgta ctccatgtaa ccaggaccag tggtccccag
     3961 agaaaagcac agcctgctta cctcgcaggc ccaagtttct ggcttggggg gagccagttg
     4021 tgctgtcact cctcctgctg ctttgcctgg tgctgggtct agcactggct gctctggggc
     4081 tctctgtcca ccactgggac agccctcttg tccaggcctc aggtggctca cagttctgct
     4141 ttggcctgat ctgcctaggc ctcttctgcc tcagtgtcct tctgttccca gggcggccaa
     4201 gctctgccag ctgccttgca caacaaccaa tggctcacct ccctctcaca ggctgcctga
     4261 gcacactctt cctgcaagca gctgagacct ttgtggagtc tgagctgcca ctgagctggg
     4321 caaactggct atgcagctac cttcggggac tctgggcctg gctagtggta ctgttggcca
     4381 cttttgtgga ggcagcacta tgtgcctggt atttgatcgc tttcccacca gaggtggtga
     4441 cagactggtc agtgctgccc acagaggtac tggagcactg ccacgtgcgt tcctgggtca
     4501 gcctgggctt ggtgcacatc accaatgcaa tgttagcttt cctctgcttt ctgggcactt
     4561 tcctggtaca gagccagcct ggccgctaca accgtgcccg tggtctcacc ttcgccatgc
     4621 tagcttattt catcacctgg gtctcttttg tgcccctcct ggccaatgtg caggtggcct
     4681 accagccagc tgtgcagatg ggtgctatcc tagtctgtgc cctgggcatc ctggtcacct
     4741 tccacctgcc caagtgctat gtgcttcttt ggctgccaaa gctcaacacc caggagttct
     4801 tcctgggaag gaatgccaag aaagcagcag atgagaacag tggcggtggt gaggcagctc
     4861 agggacacaa tgaaggatcc gactacaaag accatgacgg tgattataaa gatcatgaca
     4921 tcgactacaa ggatgacgat gacaagtagc ggccgcgaag gatctgcgat cgctccggtg
     4981 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     5041 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     5101 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     5161 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     5221 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     5281 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     5341 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     5401 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     5461 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagat gaccgagtac
     5521 aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc
     5581 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag
     5641 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5701 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5761 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5821 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5881 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5941 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     6001 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     6061 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgacaatcaa
     6121 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6181 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6241 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6301 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6361 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6421 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6481 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6541 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6601 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6661 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggtac ctttaagacc
     6721 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga
     6781 agggctaatt cactcccaac gaaaataaga tctgcttttt gcttgtactg ggtctctctg
     6841 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     6901 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     6961 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     7021 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     7081 gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     7141 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     7201 ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg
     7261 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     7321 ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag agacggccca
     7381 aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7441 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7501 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7561 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7621 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7681 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     8161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     8221 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8401 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8461 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8581 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8641 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8701 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8761 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8821 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8881 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8941 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9001 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9061 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9121 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     9181 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9241 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9301 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9361 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9421 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9481 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9541 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     9601 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     9661 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     9721 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     9781 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     9841 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     9901 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     9961 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
