LOCUS       V025682                10199 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025682
VERSION     V025682
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             3547..4818
                     /label="SKP2(NM_005983)"
                     /note="SKP2(NM_005983)"
                     /gene="SKP2"
     CDS             4822..4911
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4949..5535
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5555..6154
                     /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    6168..6756
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6963..7596
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7724..7740)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7748..7764
                     /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(7772..7802)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7817..7838
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8126..8714)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8885..9745)
                     /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(9746..9850)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9898..10032
                     /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 gatctatttc cggtgaattc
     3541 gccaccatgc acaggaagca cctccaggag attccagacc tgagtagcaa cgttgccacc
     3601 agcttcacgt ggggatggga ttccagcaag acttctgaac tgctgtcagg catgggggtc
     3661 tccgccctgg agaaagagga gcccgacagt gagaacatcc cccaggaact gctctcaaac
     3721 ctgggccacc cggagagccc cccacggaaa cggctgaaga gcaaagggag tgacaaagac
     3781 tttgtgattg tccgcaggcc taagctaaat cgagagaact ttccaggtgt ttcatgggac
     3841 tcccttccgg atgagctgct cttgggaatc ttttcctgtc tgtgcctccc tgagctgcta
     3901 aaggtctctg gtgtttgtaa gaggtggtat cgcctagcgt ctgatgagtc tctatggcag
     3961 accttagacc tcacaggtaa aaatctgcac ccggatgtga ctggtcggtt gctgtctcaa
     4021 ggggtgattg ccttccgctg cccacgatca tttatggacc aaccattggc tgaacatttc
     4081 agcccttttc gtgtacagca catggaccta tcgaactcag ttatagaagt gtccaccctc
     4141 cacggcatac tgtctcagtg ttccaagttg cagaatctaa gcctggaagg cctgcggctt
     4201 tcggatccca ttgtcaatac tctcgcaaaa aactcaaatt tagtgcgact taacctttct
     4261 gggtgttctg gattctctga atttgccctg cagactttgc taagcagctg ttccagactg
     4321 gatgagctga acctctcctg gtgttttgat ttcactgaaa agcatgtaca ggtggctgtt
     4381 gcgcatgtgt cagagaccat cacccagctg aatcttagcg gctacagaaa gaatctccag
     4441 aaatcagatc tctctacttt agttagaaga tgccccaatc ttgtccatct agacttaagt
     4501 gatagtgtca tgctaaagaa tgactgcttt caggaatttt tccagctcaa ctacctccaa
     4561 cacctatcac tcagtcggtg ctatgatata atacctgaaa ctttacttga acttggagaa
     4621 attcccacac taaaaacact acaagttttt ggaatcgtgc cagatggtac ccttcaactg
     4681 ttaaaggaag cccttcctca tctacagatt aattgctccc atttcaccac cattgccagg
     4741 ccaactattg gcaacaaaaa gaaccaggag atatggggca tcaaatgccg actgacactg
     4801 caaaagccca gttgtctatt cgaacaaaaa ctaatatcag aggaagattt ggaacagaaa
     4861 ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt atagctcgag
     4921 actagttcta gagcggccgc ggatcccgcc cctctccctc ccccccccct aacgttactg
     4981 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     5041 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     5101 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     5161 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     5221 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     5281 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     5341 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     5401 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     5461 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     5521 aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg
     5581 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     5641 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     5701 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     5761 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     5821 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     5881 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     5941 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     6001 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     6061 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     6121 cacctggtgc atgacccgca agcccggtgc ctagacgcgt ctggaacaat caacctctgg
     6181 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     6241 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     6301 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     6361 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     6421 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     6481 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     6541 attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca
     6601 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     6661 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     6721 agacgagtcg gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga
     6781 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
     6841 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
     6901 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg
     6961 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
     7021 cacacaaggc tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc
     7081 actgaccttt ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc
     7141 caataaagga gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
     7201 ggagagagaa gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg
     7261 agagctgcat ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc
     7321 cgctggggac tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag
     7381 atcctgcata taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct
     7441 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     7501 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     7561 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     7621 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct
     7681 agcgtttacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     7741 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     7801 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     7861 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     7921 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     7981 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     8041 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     8101 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     8161 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     8221 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     8281 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     8341 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     8401 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     8461 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     8521 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     8581 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     8641 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     8701 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     8761 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     8821 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     8881 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     8941 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     9001 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     9061 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     9121 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     9181 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     9241 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     9301 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     9361 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     9421 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     9481 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     9541 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     9601 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     9661 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     9721 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     9781 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     9841 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac
     9901 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     9961 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
    10021 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
    10081 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
    10141 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
