LOCUS       V026005                10075 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026005
VERSION     V026005
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"
     CDS             2843..4771
                     /label="CLINT1(NM_001195555)"
                     /note="CLINT1(NM_001195555)"
                     /gene="CLINT1"
     CDS             4778..4843
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4863..5362
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5383..5982
                     /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    5996..6584
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6791..7424
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7553..7569)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7577..7593
                     /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(7601..7631)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7646..7667
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7955..8543)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8714..9574)
                     /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(9575..9679)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9727..9861
                     /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 ctcaagcttc gaattcgcca ccatgttgaa catgtggaag gtgcgcgagc tggtggacaa
     2881 agccaccaat gttgttatga attattcaga gatcgagtct aaggttcgag aggcaacgaa
     2941 cgatgatcct tggggacctt ctgggcaact catgggagag attgccaagg ctacatttat
     3001 gtatgaacaa tttccagaac ttatgaacat gctttggtca cgaatgttaa aagacaacaa
     3061 aaagaattgg agaagagttt ataagtcgtt gctgctccta gcttacctca taaggaatgg
     3121 atcagagcgt gttgttacaa gtgccagaga acacatttat gatttacgat ccctggaaaa
     3181 ttaccacttt gtagatgagc atggtaagga tcaaggtata aatattcgac agaaggtgaa
     3241 ggaattggtt gaatttgccc aggatgacga caggcttcgt gaagagcgaa agaaagcaaa
     3301 gaagaacaaa gacaagtatg ttggggtttc ctcagacagt gttggaggat tcagatacag
     3361 tgaaagatat gatcctgagc ccaaatcaaa atgggatgag gagtgggata aaaacaagag
     3421 tgcttttcca ttcagtgata aattaggtga gctgagtgat aaaattggaa gcacaattga
     3481 tgacaccatc agcaagttcc ggaggaaaga tagagaagac tctccagaaa gatgcagcga
     3541 cagcgatgag gaaaagaaag cgagaagagg cagatctccc aaaggtgaat tcaaagatga
     3601 agaggagact gtgacgacaa agcatattca tatcacacag gccacagaga ccaccacaac
     3661 cagacacaag cgcacagcaa atccttccaa aaccattgat cttggagcag cagcacatta
     3721 cacaggggac aaagcaagtc cagatcagaa tgcttcaacc cacacacctc agtcttcagt
     3781 taagacttca gtgcctagca gcaagtcatc tggtgacctt gttgatctgt ttgatggcac
     3841 cagccagtca acaggaggat cagctgattt attcggagga tttgctgact ttggctcagc
     3901 tgctgcatca ggcagtttcc cttcccaagt aacagcaaca agtgggaatg gagactttgg
     3961 tgactggagt gccttcaacc aagccccatc aggccctgtt gcttccagtg gcgagttctt
     4021 tggcagtgcc tcacagccag cggtagaact tgttagtggc tcacaatcag ctctaggccc
     4081 acctcctgct gcctcaaatt cttcagacct gtttgatctt atgggctcgt cccaggcaac
     4141 catgacatct tcccagagta tgaatttctc tatgatgagc actaacactg tgggacttgg
     4201 tttgcctatg tcaagatcac agcctttgca aaatgttagc acagtgctgc agaagcctaa
     4261 tcctctctat aatcagaata cagatatggt ccagaaatca gtcagcaaaa ccttgccctc
     4321 tacttggtct gaccccagtg taaacatcag cctagacaac ttactacctg gtatgcagcc
     4381 ttccaaaccc cagcagccat cactgaatac aatgattcag caacagaata tgcagcagcc
     4441 tatgaatgtg atgactcaaa gttttggagc tgtgaacctc agttctccat cgaacatgct
     4501 tcctgtccgg ccccaaacta atgctttgat agggggaccc atgcctatga gcatgcccaa
     4561 tgtgatgact ggcaccatgg gaatggcccc tcttggaaat actccgatga tgaaccagag
     4621 catgatgggc atgaacatga acatagggat gtccgctgct gggatgggct tgacaggcac
     4681 aatgggaatg ggcatgccca acatagccat gacttctgga actgtgcaac ccaagcaaga
     4741 tgcctttgca aatttcgcca attttagcaa aggatccgac tacaaagacc atgacggtga
     4801 ttataaagat catgacatcg actacaagga tgacgatgac aagtagtcta gataattcta
     4861 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
     4921 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
     4981 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
     5041 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
     5101 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
     5161 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
     5221 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
     5281 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
     5341 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc
     5401 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     5461 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     5521 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     5581 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     5641 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     5701 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     5761 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     5821 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     5881 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     5941 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca
     6001 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6061 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6121 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6181 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6241 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6301 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6361 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg
     6421 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6481 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6541 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc
     6601 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct
     6661 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag
     6721 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     6781 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg
     6841 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     6901 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     6961 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     7021 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     7081 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     7141 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     7201 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     7261 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     7321 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     7381 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     7441 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg
     7501 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     7561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     7621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     7681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     7741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     7801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     7861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     7921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     7981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     8041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     8101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     8161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     8221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     8281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     8341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     8401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     8461 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     8521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     8581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     8641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     8701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     8761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     8821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     8881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     8941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     9001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     9061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     9121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     9181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     9241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     9301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     9361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     9421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     9481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     9541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     9601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     9661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     9721 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     9781 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     9841 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
     9901 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
     9961 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
    10021 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
