LOCUS       V042069                10159 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042069
VERSION     V042069
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"
     promoter        2337..3519
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3548..4825
                     /label="STK25(NM_001271977)"
                     /note="STK25(NM_001271977)"
                     /gene="STK25"
     CDS             4832..4897
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4909..5495
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5515..6114
                     /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    6128..6716
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6923..7556
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7684..7700)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7708..7724
                     /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(7732..7762)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7777..7798
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8086..8674)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8845..9705)
                     /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(9706..9810)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9858..9992
                     /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt
     3541 cgccaccatg gctcacctcc ggggatttgc caaccagcac tctcgagtgg accctgagga
     3601 gctcttcacc aagctcgacc gcattggcaa gggctcgttt ggggaggtct acaagggcat
     3661 cgataaccac acaaaggagg tggtggccat caagatcatc gacctggagg aggccgagga
     3721 tgagatcgag gacatccagc aggagatcac tgtcctcagt cagtgcgaca gcccctacat
     3781 cacccgctac tttggctcct acctaaagag caccaagcta tggatcatca tggagtacct
     3841 gggcggcggc tcagcactgg acttgcttaa accaggtccc ctggaggaga catacattgc
     3901 cacgatcctg cgggagattc tgaagggcct ggattatctg cactccgaac gcaagatcca
     3961 ccgagacatc aaagctgcca acgtgctact ctcggagcag ggtgacgtga agctggcgga
     4021 ctttggggta gcagggcagc tcacagacac gcagattaag aggaacacat tcgtgggcac
     4081 ccccttctgg atggcacctg aggtcatcaa gcagtcggcc tacgacttca aggctgacat
     4141 ctggtccctg gggatcacag ccatcgagct ggccaagggg gagcctccaa actctgacct
     4201 ccaccccatg cgcgtcctgt tcctgattcc caagaacagc ccacccacac tggagggcca
     4261 gcacagcaag cccttcaagg agttcgtgga ggcctgcctc aacaaagacc cccgattccg
     4321 gcccacggcc aaggagctcc tgaagcacaa gttcatcaca cgctacacca agaagacctc
     4381 cttcctcacg gagctcatcg accgctataa gcgctggaag tcagaggggc atggcgagga
     4441 gtccagctct gaggactctg acattgatgg cgaggcggag gacggggagc agggccccat
     4501 ctggacgttc ccccctacca tccggccgag tccacacagc aagcttcaca aggggacggc
     4561 cctgcacagt tcacagaagc ctgcggagcc cgtcaagagg cagccgaggt cccagtgcct
     4621 gtccacgctg gtccggcccg tcttcggaga gctcaaagag aagcacaagc agagcggcgg
     4681 gagcgtgggt gcgctggagg agctggagaa cgccttcagc ctggccgagg agtcctgccc
     4741 cggcatctca gacaagctga tggtgcacct ggtggagcga gtgcagaggt tttcacacaa
     4801 cagaaaccac ctgacatcca cccgcggccg cgactacaaa gaccatgacg gtgattataa
     4861 agatcatgac atcgactaca aggatgacga tgacaagtag tctagacgcc cctctccctc
     4921 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     4981 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     5041 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     5101 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     5161 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     5221 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     5281 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     5341 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     5401 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     5461 cctttgaaaa acacgatgat aagcttgcca caacccacaa ggagacgacc ttccatgacc
     5521 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     5581 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     5641 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5701 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5761 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5821 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5881 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5941 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     6001 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     6061 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctagacgcgt
     6121 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     6181 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     6241 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     6301 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     6361 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     6421 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6481 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     6541 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6601 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6661 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6721 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     6781 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     6841 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6901 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc
     6961 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     7021 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     7081 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     7141 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     7201 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     7261 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     7321 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     7381 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     7441 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7501 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     7561 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7621 tgagaggcct tgacattgct agcgtttacc gtcgacctct agctagagct tggcgtaatc
     7681 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     7801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     8041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     8101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     8161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     8221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     8281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     8341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     8401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     8461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     8521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     9001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     9061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     9121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     9181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     9241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     9301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     9361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     9421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     9481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9841 cgacggatcg ggagatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     9901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     9961 aactcatcaa tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc
    10021 atcccaaact tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac
    10081 tctaaacctg tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat
    10141 gtactacaaa cttagtagt
//
