LOCUS       V027743                 9727 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027743
VERSION     V027743
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..4423
                     /label="RAG2(NM_000536)"
                     /note="RAG2(NM_000536)"
                     /gene="RAG2"
     CDS             4430..4495
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4515..5014
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5035..5634
                     /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    5648..6236
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6443..7076
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7205..7221)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7229..7245
                     /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(7253..7283)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7298..7319
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7607..8195)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8366..9226)
                     /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(9227..9331)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9379..9513
                     /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 ccatgtctct gcagatggta acagtcagta ataacatagc
     2881 cttaattcag ccaggcttct cactgatgaa ttttgatgga caagttttct tctttggaca
     2941 aaaaggctgg cccaaaagat cctgccccac tggagttttc catctggatg taaagcataa
     3001 ccatgtcaaa ctgaagccta caattttctc taaggattcc tgctacctcc ctcctcttcg
     3061 ctacccagcc acttgcacat tcaaaggcag cttggagtct gaaaagcatc aatacatcat
     3121 ccatggaggg aaaacaccaa acaatgaggt ttcagataag atttatgtca tgtctattgt
     3181 ttgcaagaac aacaaaaagg ttacttttcg ctgcacagag aaagacttgg taggagatgt
     3241 tcctgaagcc agatatggtc attccattaa tgtggtgtac agccgaggga aaagtatggg
     3301 tgttctcttt ggaggacgct catacatgcc ttctacccac agaaccacag aaaaatggaa
     3361 tagtgtagct gactgcctgc cctgtgtttt cctggtggat tttgaatttg ggtgtgctac
     3421 atcatacatt cttccagaac ttcaggatgg gctatctttt catgtctcta ttgccaaaaa
     3481 tgacaccatc tatattttag gaggacattc acttgccaat aatatccggc ctgccaacct
     3541 gtacagaata agggttgatc ttcccctggg tagcccagct gtgaattgca cagtcttgcc
     3601 aggaggaatc tctgtctcca gtgcaatcct gactcaaact aacaatgatg aatttgttat
     3661 tgttggtggc tatcagcttg aaaatcaaaa aagaatgatc tgcaacatca tctctttaga
     3721 ggacaacaag atagaaattc gtgagatgga gaccccagat tggaccccag acattaagca
     3781 cagcaagata tggtttggaa gcaacatggg aaatggaact gtttttcttg gcataccagg
     3841 agacaataaa caagttgttt cagaaggatt ctatttctat atgttgaaat gtgctgaaga
     3901 tgatactaat gaagagcaga caacattcac aaacagtcaa acatcaacag aagatccagg
     3961 ggattccact ccctttgaag actctgaaga attttgtttc agtgcagaag caaatagttt
     4021 tgatggtgat gatgaatttg acacctataa tgaagatgat gaagaagatg agtctgagac
     4081 aggctactgg attacatgct gccctacttg tgatgtggat atcaacactt gggtaccatt
     4141 ctattcaact gagctcaaca aacccgccat gatctactgc tctcatgggg atgggcactg
     4201 ggtccatgct cagtgcatgg atctggcaga acgcacactc atccatctgt cagcaggaag
     4261 caacaagtat tactgcaatg agcatgtgga gatagcaaga gctctacaca ctccccaaag
     4321 agtcctaccc ttaaaaaagc ctccaatgaa atccctccgt aaaaaaggtt ctggaaaaat
     4381 cttgactcct gccaagaaat cctttcttag aaggttgttt gatggatccg actacaaaga
     4441 ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg acaagtagtc
     4501 tagataattc taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt
     4561 agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac
     4621 atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac
     4681 tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca
     4741 aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa
     4801 gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca
     4861 gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg
     4921 cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc
     4981 gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcccaagct taccatgacc
     5041 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5101 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     5161 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5221 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5281 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5341 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5401 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5461 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     5521 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5581 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgaccgcgt
     5641 ctggaacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     5701 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     5761 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     5821 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     5881 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     5941 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     6001 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     6061 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     6121 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     6181 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     6241 aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca
     6301 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca
     6361 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     6421 tttttaaaag aaaagagggg actggaaggg ctaattcact cccaacgaag acaagatatc
     6481 cttgatctgt ggatctacca cacacaaggc tacttccctg attagcagaa ctacacacca
     6541 gggccagggg tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag
     6601 ccagataagg tagaagaggc caataaagga gagaacacca gcttgttaca ccctgtgagc
     6661 ctgcatggga tggatgaccc ggagagagaa gtgttagagt ggaggtttga cagccgccta
     6721 gcatttcatc acgtggcccg agagctgcat ccggagtact tcaagaactg ctgatatcga
     6781 gcttgctaca agggactttc cgctggggac tttccaggga ggcgtggcct gggcgggact
     6841 ggggagtggc gagccctcag atcctgcata taagcagctg ctttttgcct gtactgggtc
     6901 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     6961 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     7021 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     7081 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     7141 tgagaggcct tgacattgct agcgttttac cgtcgacctc tagctagagc ttggcgtaat
     7201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     7261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     7321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     7381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     7441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     7501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     7561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     8701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     9001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9361 tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     9421 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     9481 aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat
     9541 catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta
     9601 ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata
     9661 tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt actacaaact
     9721 tagtagt
//
