LOCUS       V033900                 9825 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033900
VERSION     V033900
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..4531
                     /label="ITPRIPL1(NM_178495)"
                     /note="ITPRIPL1(NM_178495)"
                     /gene="ITPRIPL1"
     CDS             4532..4555
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             4559..4582
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4613..5112
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5133..5732
                     /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    5746..6334
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6541..7174
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7303..7319)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7327..7343
                     /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(7351..7381)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7396..7417
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7705..8293)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8464..9324)
                     /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(9325..9429)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9477..9611
                     /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 ccatgaccac tgactcctac tctcctactt ctccagatgc
     2881 agaggcctcc atggctgtga taagcctgct gttcttggca gtgatgtatg ttgttcacca
     2941 ccctctgatg gtcagtgatc ggatggacct ggacacatta gccaggagtc ggcagctgga
     3001 gaagcgaatg agtgaggaga tgcgcctgct agagatggag tttgaagaga gaaagcgagc
     3061 cgctgagcag aggcagaagg cagagaactt ctggacagga gacacatcca gtgaccagtt
     3121 agtgctgggg aagaaagaca tggggtggcc gttccaggcc gatggccagg aagggcctct
     3181 gggctggatg ctgggaaacc tgtggaacac tggcctcttt tgcctttttc tcgtctttga
     3241 gctcctgcga cagaacatgc agcatgaacc ggcctttgat tccagcagtg aggaggagga
     3301 ggaggaagtc cgtgttgtcc ctgtcacctc ttacaactgg cttactgact tcccctccca
     3361 ggaggccctg gactcctttt acaaacacta tgtccaaaat gccatccgtg acctgccctg
     3421 cacctgtgag tttgtggaga gttttgtgga tgatctcatt gaggcctgtc gggtgctcag
     3481 ccgccaagag gctcacccac aattggaaga ctgcctgggc atcggggctg cctttgagaa
     3541 atggggaacc ctccatgaga cccagaaatt tgatatcctg gtgcccattg tccccccaca
     3601 gggcaccatg tttgtcctgg agatgaggga cccagccctg ggccgccgct gtggctgtgt
     3661 gctggtggag tcagaatgtg tgtgcaagcg tgagaaactc ctaggggacg tgctgtgcct
     3721 ggtgcaccac cacagggacc cctcggcagt cttggggaag tgtagtagct ccatcaaggc
     3781 agctctctgc accggcttcc acctagacgt gtgcaagact gtgcagtggt tccggaacat
     3841 gatgggcaat gcctgggccc ttgtggccca caagtatgac tttaaactca gtctcccacc
     3901 gtctaccacc tcctgcaagc tccggctgga ctatcgctca ggccgctttc tctcaatcca
     3961 cttggtcctg ggggtgcaac gagaagacac cttggtctac ctggtgagtc aggctcctga
     4021 ccaggagcag ctcaccagtg tggactggcc tgagtccttt gtggcctgtg agcacctgtt
     4081 cctgaagctg gtggggcgct ttgcccccga gaacacctgt cacctcaagt gcctccagat
     4141 cattttaagt ctccggcagc atcagagctt accccatgga gcatcccgcc ccatcctcac
     4201 ttcttaccat tttaaaacag ctctcatgca cctcttgcta cggctgcccc tcacggactg
     4261 ggcccacaac atgctctctc agcggctcca ggacattctc tggttcttgg gccgtggcct
     4321 ccagcaaagg tccctccatc atttcctcat tggtaacaat tttctgcccc tgaccatccc
     4381 aatccctaag acatttagga atgctgagcc ggtcaatctc ttccaacacc tggtgctcaa
     4441 ccccaaggca cattcacagg cagtggaaga gttccaaaac cttctgaccc aggtgaaaac
     4501 tctgcctcat gccccactgg ctgcagcacc tgactacaag gacgacgatg acaaggcaga
     4561 ctacaaggac gacgatgaca agtaggatcc cgcgactcta gataattcta ccgggtaggg
     4621 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
     4681 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
     4741 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
     4801 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
     4861 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
     4921 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
     4981 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
     5041 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
     5101 ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc acggtgcgcc
     5161 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     5221 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     5281 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     5341 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     5401 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     5461 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     5521 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     5581 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     5641 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     5701 cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca acctctggat
     5761 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     5821 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     5881 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     5941 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     6001 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     6061 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     6121 tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc
     6181 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     6241 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     6301 acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc
     6361 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     6421 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     6481 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aagaggggac
     6541 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     6601 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
     6661 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
     6721 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     6781 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
     6841 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
     6901 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
     6961 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
     7021 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     7081 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7141 agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca tcttattatt
     7201 cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg acattgctag
     7261 cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     7321 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7381 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7441 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7501 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7561 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7621 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7681 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7741 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7801 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7861 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7921 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7981 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     8041 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     8101 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     8161 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     8221 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     8281 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     8341 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8401 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8461 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8521 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8581 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8641 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8701 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8761 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8821 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8881 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8941 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     9001 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     9061 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     9121 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9181 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9241 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9301 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9361 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9421 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatcaact
     9481 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     9541 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9601 atgtctggat caactggata actcaagcta accaaaatca tcccaaactt cccaccccat
     9661 accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt gattcctctg
     9721 aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac ttagtagttt
     9781 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
