LOCUS       V020244                 8822 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020244
VERSION     V020244
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             2290..2928
                     /label="VHL(NM_000551)"
                     /note="VHL(NM_000551)"
                     /gene="VHL"
     CDS             2929..2994
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3043..3254
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3267..3535
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     CDS             3570..4262
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="red fluorescent protein from Entacmaea
                     quadricolor"
                     /transl_table=1
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
                     /label="TurboRFP"
     CDS             4263..4316
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4317..4916
                     /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    4917..5505
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5579..5812
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5884..6005
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6024..6159
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6197..6213)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6221..6237
                     /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(6245..6275)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6290..6311
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6599..7187)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7358..8218)
                     /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(8219..8323)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8797..8813
                     /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 agagccacca tgccccggag ggcggagaac tgggacgagg ccgaggtagg cgcggaggag
     2341 gcaggcgtcg aagagtacgg ccctgaagaa gacggcgggg aggagtcggg cgccgaggag
     2401 tccggcccgg aagagtccgg cccggaggaa ctgggcgccg aggaggagat ggaggccggg
     2461 cggccgcggc ccgtgctgcg ctcggtgaac tcgcgcgagc cctcccaggt catcttctgc
     2521 aatcgcagtc cgcgcgtcgt gctgcccgta tggctcaact tcgacggcga gccgcagccc
     2581 tacccaacgc tgccgcctgg cacgggccgc cgcatccaca gctaccgagg tcacctttgg
     2641 ctcttcagag atgcagggac acacgatggg cttctggtta accaaactga attatttgtg
     2701 ccatctctca atgttgacgg acagcctatt tttgccaata tcacactgcc agtgtatact
     2761 ctgaaagagc gatgcctcca ggttgtccgg agcctagtca agcctgagaa ttacaggaga
     2821 ctggacatcg tcaggtcgct ctacgaagat ctggaagacc acccaaatgt gcagaaagac
     2881 ctggagcggc tgacacagga gcgcattgca catcaacgga tgggagatga ctacaaagac
     2941 catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga caagtaggat
     3001 ccgcggccgc aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc
     3061 ccacagtccc cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt
     3121 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
     3181 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
     3241 ccgccagaac acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc
     3301 ctacctgagg ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg
     3361 cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt
     3421 gtccggcgct cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc
     3481 tgcttgctca actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc
     3541 tgtgaccggc gcctacgcta gacgccacca tgagcgagct gatcaaggag aacatgcaca
     3601 tgaagctgta catggagggc accgtgaaca accaccactt caagtgcaca tccgagggcg
     3661 aaggcaagcc ctacgagggc acccagacca tgaagatcaa ggtggtcgag ggcggccctc
     3721 tccccttcgc cttcgacatc ctggctacca gcttcatgta cggcagcaaa gccttcatca
     3781 accacaccca gggcatcccc gacttcttta agcagtcctt ccctgagggc ttcacatggg
     3841 agagaatcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcttcc
     3901 agaacggctg catcatctac aacgtcaaga tcaacggggt gaacttccca tccaacggcc
     3961 ctgtgatgca gaagaaaaca cgcggctggg aggccaacac cgagatgctg taccccgctg
     4021 acggcggcct gagaggccac agccagatgg ccctgaagct cgtgggcggg ggctacctgc
     4081 actgctcctt caagaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg
     4141 gcttccactt cgtggaccac agactggaaa gaatcaagga ggccgacaaa gagacctacg
     4201 tcgagcagca cgagatggct gtggccaagt actgcgacct ccctagcaaa ctggggcaca
     4261 gagagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatga
     4321 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4381 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4441 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4501 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4561 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4621 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4681 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4741 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4801 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4861 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaaatc
     4921 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4981 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     5041 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     5101 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     5161 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     5221 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     5281 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     5341 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5401 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5461 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctggt acctttaaga
     5521 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
     5581 gaagggctaa ttcactccca acgaaaataa gatctgcttt ttgcttgtac tgggtctctc
     5641 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5701 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5761 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtagtagt
     5821 tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag
     5881 aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5941 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6001 tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccagttcc gcccattctc
     6061 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     6121 agctattcca gaagtagtga ggaggctttt ttggaggcct agacttttgc agagacggcc
     6181 caaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7021 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8341 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     8401 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     8461 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     8521 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     8581 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     8641 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     8701 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     8761 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc
     8821 tg
//
