LOCUS       V038866                 9947 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038866
VERSION     V038866
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     regulatory      2820..2829
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             2829..2894
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             2901..4703
                     /label="DYRK2(NM_006482)"
                     /note="DYRK2(NM_006482)"
                     /gene="DYRK2"
     promoter        4735..5234
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5255..5854
                     /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    5868..6456
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6663..7296
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7425..7441)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7449..7465
                     /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(7473..7503)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7518..7539
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7827..8415)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8586..9446)
                     /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(9447..9551)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9599..9733
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
     2881 atgacgatga caaggaattc atgttaacca ggaaaccttc ggccgccgct cccgccgcct
     2941 acccgaccgg ccgaggtggg gacagcgccg ttcgtcagct tcaggcttcc ccggggctcg
     3001 gtgcaggggc cacccggagc ggagtgggga ctggcccgcc ctcccccatc gccctgccgc
     3061 ctctccgggc cagcaacgct gccgccgcag cccacacgat tggcggcagt aagcacacaa
     3121 tgaatgatca cctgcatgtc ggcagccacg ctcacggaca gatccaggtt caacagttgt
     3181 ttgaggataa cagtaacaag cggacagtgc tcacgacaca accaaatggg cttacaacag
     3241 tgggcaaaac gggcttgcca gtggtgccag agcggcagct ggacagcatt catagacggc
     3301 aggggagctc cacctctcta aagtccatgg aaggcatggg gaaggtgaaa gccaccccca
     3361 tgacacctga acaagcaatg aagcaataca tgcaaaaact cacagccttc gaacaccatg
     3421 agattttcag ctaccctgaa atatatttct tgggtctaaa tgctaagaag cgccagggca
     3481 tgacaggtgg gcccaacaat ggtggctatg atgatgacca gggatcatat gtgcaggtgc
     3541 cccacgatca cgtggcttac aggtatgagg tcctcaaggt cattgggaag gggagctttg
     3601 ggcaggtggt caaggcctac gatcacaaag tccaccagca cgtggcccta aagatggtgc
     3661 ggaatgagaa gcgcttccac cggcaagcag cggaggagat ccgaatcctg gaacacctgc
     3721 ggaagcagga caaggataac acaatgaatg tcatccatat gctggagaat ttcaccttcc
     3781 gcaaccacat ctgcatgacg tttgagctgc tgagcatgaa cctctatgag ctcatcaaga
     3841 agaataaatt ccagggcttc agtctgcctt tggttcgcaa gtttgcccac tcgattctgc
     3901 agtgcttgga tgctttgcac aaaaacagaa taattcactg tgaccttaag cccgagaaca
     3961 ttttgttaaa gcagcagggt agaagcggta ttaaagtaat tgattttggc tccagttgtt
     4021 acgagcatca gcgtgtctac acgtacatcc agtcgcgttt ttaccgggct ccagaagtga
     4081 tccttggggc caggtatggc atgcccattg atatgtggag cctgggctgc attttagcag
     4141 agctcctgac gggttacccc ctcttgcctg gggaagatga aggggaccag ctggcctgta
     4201 tgattgaact gttgggcatg ccctcacaga aactgctgga tgcatccaaa cgagccaaaa
     4261 attttgtgag ctccaagggt tatccccgtt actgcactgt cacgactctc tcagatggct
     4321 ctgtggtcct aaacggaggc cgttcccgga gggggaaact gaggggccca ccggagagca
     4381 gagagtgggg gaacgcgctg aaggggtgtg atgatcccct tttccttgac ttcttaaaac
     4441 agtgtttaga gtgggatcct gcagtgcgca tgaccccagg ccaggctttg cggcacccct
     4501 ggctgaggag gcggttgcca aagcctccca ccggggagaa aacgtcagtg aaaaggataa
     4561 ctgagagcac cggtgctatc acatctatat ccaagttacc tccaccttct agctcagctt
     4621 ccaaactgag gactaatttg gcgcagatga cagatgccaa tgggaatatt cagcagagga
     4681 cagtgttgcc aaaacttgtt agctgaggat cccgcgactc tagataattc taccgggtag
     4741 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     4801 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     4861 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     4921 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     4981 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     5041 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     5101 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     5161 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     5221 ctccgggcct ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg
     5281 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     5341 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     5401 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5461 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5521 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5581 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5641 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     5701 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     5761 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     5821 cacctggtgc atgacccgca agcccggtgc ctgaccgcgt ctggaacaat caacctctgg
     5881 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5941 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6001 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6061 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6121 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6181 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6241 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6301 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6361 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6421 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     6481 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6541 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     6601 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6661 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     6721 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     6781 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     6841 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     6901 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     6961 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     7021 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     7081 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     7141 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7201 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7261 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     7321 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     7381 agcgttttac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     7441 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7501 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     7561 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7621 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7681 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7741 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7801 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7861 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7921 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7981 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8041 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8101 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8161 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8221 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8281 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8341 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8401 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8461 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8521 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8581 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8641 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8701 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8761 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8821 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8881 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8941 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     9001 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9061 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9121 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9181 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     9241 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     9301 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     9361 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     9421 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     9481 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     9541 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa
     9601 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     9661 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     9721 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     9781 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     9841 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     9901 ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
