LOCUS       V039334                10312 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039334
VERSION     V039334
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..4999
                     /label="HOOK2(NM_013312)"
                     /note="HOOK2(NM_013312)"
                     /gene="HOOK2"
     CDS             5015..5080
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        5100..5599
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5620..6219
                     /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    6233..6821
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7028..7661
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7790..7806)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7814..7830
                     /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(7838..7868)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7883..7904
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8192..8780)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8951..9811)
                     /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(9812..9916)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9964..10098
                     /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 ccatgagcgt ggacaaagct gagctatgcg ggtctctgct
     2881 cacctggtta cagacgttcc acgttccgtc tccctgtgcc agccctcagg acctgagcag
     2941 cggccttgcc gtagcctatg tgctgaacca gatagacccc tcctggttca acgaggcatg
     3001 gctccagggc atctcggaag atccaggtcc caactggaag ctgaaggtca gcaatctgaa
     3061 gatggtctta cggagcctag tagagtactc ccaggatgtc ctggcgcatc ctgtgtcaga
     3121 agagcatctc ccagatgtga gcctcattgg agagttctca gacccggcag agctcggcaa
     3181 gctgcttcag ctggtgctgg gctgtgccat cagttgcgag aaaaagcagg accacatcca
     3241 gagaatcatg acgctggaag aatcggttca gcatgtggtg atggaagcca tccaagagct
     3301 catgaccaaa gacactcctg actccctgtc accagagacg tatggcaact ttgacagcca
     3361 gtcccgcagg tactatttcc taagtgagga ggctgaggag ggggacgaat tacagcagcg
     3421 ctgtctggat ctggagcggc agctgatgct cctgtcagag gagaagcaga gcctggcgca
     3481 agagaatgca gggctgcggg agcggatggg ccggcctgaa ggcgagggta ccccaggtct
     3541 cactgccaag aagctgctgc tgctgcaatc ccagctggag cagttgcagg aggagaactt
     3601 caggctggag agtggcaggg aggatgagcg cctgcgctgt gccgagctgg agagggaggt
     3661 tgcggagctg cagcaccgga accaggcgct gactagcctg gcccaggagg cacaggccct
     3721 gaaggatgag atggatgaac tacggcagtc ttcggagcgt gctgggcagc tggaggccac
     3781 gctgaccagt tgccggcgcc gcttgggcga gctgagggag ctgcggcggc aggtgcggca
     3841 gctggaggaa cgcaacgccg gccacgccga gcgcacgcga caactggagg atgagctacg
     3901 ccgagcgggc tccctgcgcg cccagctgga ggcgcagcgg cggcaggtgc aggaactgca
     3961 gggccagcgg caggaggagg ccatgaaggc cgagaaatgg ctatttgaat gccgcaacct
     4021 ggaggaaaag tatgagtcgg tgacaaagga gaaggagcgg ctgttggcgg agcgggactc
     4081 cttgcgggag gccaatgagg agctgcgctg cgcccagctg cagccgcggg ggttgaccca
     4141 ggccgatccc tcactggatc ccacctccac acccgtggat aacttagccg cagagatcct
     4201 gcctgcggag ctcagggaga cgctcctgcg gcttcagctg gagaacaagc ggctgtgcag
     4261 gcaggaggcg gccgaccggg agcggcagga ggagctgcag cgccacctgg aggatgccaa
     4321 ccgcgcgcgc cacgggttgg agacgcagca ccggctgaac cagcagcagc tatccgagct
     4381 gcgggcccag gtggaggacc tgcagaaagc cctgcaggag caggggggca agactgaaga
     4441 tgccatttcc attttgctga aaaggaagct ggaggaacat ttgcagaagc ttcatgaggc
     4501 agatctggag ttgcagagga agcgggagta cattgaggag ctggagccac ccactgacag
     4561 cagcacagcc cggcggatcg aggagctgca gcataacttg cagaagaagg acgcggactt
     4621 gcgggccatg gaggagcgat accgccgcta cgtggacaag gcccgcatgg tcatgcagac
     4681 catggaaccc aagcagcggc cagctgcggg ggcacctcca gaactccatt ccctgaggac
     4741 acagctccga gaacgggatg tccgcatccg acacctggag atggactttg agaaaagccg
     4801 aagtcagcgg gagcaggaag aaaagctgct catcagtgcc tggtataata tgggcatggc
     4861 cttgcagcag cgagctgggg aggagcgggc gcctgcccat gcccagtcat tcctggcaca
     4921 gcagcggctg gcaaccaatt ctcgccgtgg acccttggga cgcctggcat ctctgaacct
     4981 tcgccccact gacaagcacg gtggaggcgg atccgactac aaagaccatg acggtgatta
     5041 taaagatcat gacatcgact acaaggatga cgatgacaag tagtctagat aattctaccg
     5101 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     5161 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     5221 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     5281 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     5341 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     5401 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     5461 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     5521 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     5581 ctcatctccg ggcctttcga cctgcagccc aagcttacca tgaccgagta caagcccacg
     5641 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     5701 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     5761 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5821 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5881 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5941 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     6001 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     6061 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     6121 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     6181 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc
     6241 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     6301 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     6361 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     6421 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     6481 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     6541 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     6601 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     6661 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     6721 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     6781 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta attctgcagt
     6841 cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac caatgctgat
     6901 tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac acctcaggta
     6961 cctttaagac caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag
     7021 aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga tctgtggatc
     7081 taccacacac aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga
     7141 tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa
     7201 gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat
     7261 gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg
     7321 gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga
     7381 ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc
     7441 ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca
     7501 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
     7561 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
     7621 atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt catgtcatct
     7681 tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga ggccttgaca
     7741 ttgctagcgt tttaccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     7801 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     7861 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     7921 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     7981 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     8041 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     8101 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     8161 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     8221 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     8281 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     8341 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     8401 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     8461 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     8521 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     8581 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     8641 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     8701 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     8761 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     8821 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     8881 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     8941 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     9001 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     9061 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     9121 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     9181 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     9241 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     9301 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     9361 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     9421 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     9481 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     9541 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     9601 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     9661 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     9721 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     9781 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     9841 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     9901 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     9961 atcaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
    10021 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
    10081 tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc caaacttccc
    10141 accccatacc ctattaccac tgccaattac ctgtggtttc atttactcta aacctgtgat
    10201 tcctctgaat tattttcatt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta
    10261 gtagttttta aagaaattgt atttgttaaa tatgtactac aaacttagta gt
//
