LOCUS       V034651                10441 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034651
VERSION     V034651
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             2815..4932
                     /label="Mre11a(NM_018736)"
                     /note="Mre11a(NM_018736)"
                     /gene="Mre11a"
     CDS             4939..5004
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    5015..5601
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5602..6396
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     misc_feature    6410..6998
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7205..7838
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7966..7982)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7990..8006
                     /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(8014..8044)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8059..8080
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8368..8956)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9127..9987)
                     /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(9988..10092)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10140..10274
                     /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 tctatttccg gtgaattcgc caccatgagc
     2821 cccacagatc cacttgacga tgaagacaca tttaaaatcc tggttgccac tgatattcat
     2881 cttggattta tggagaaaga tgcagttaga ggaaatgata catttgtgac atttgatgaa
     2941 atcttaagac ttgccctgga aaatgaagtg gattttattt tgttaggtgg tgatcttttc
     3001 catgaaaaca agccctcgag gaaaaccctg cacagctgct tggagctgct taggaagtac
     3061 tgtatgggtg accgccctgt gcagtttgag gtcatcagtg atcagtcagt caactttggt
     3121 tttagtaagt ttccatgggt gaactaccag gatggcaatc tcaacatttc cattccagta
     3181 tttagtatcc acggcaacca tgatgatccc acgggggcag atgccctctg tgccctggat
     3241 gttttaagct gtgctgggtt tgtgaatcac tttggacggt caatgtcggt ggagaaggtt
     3301 gacattagtc cggttctgct gcagaaagga agcacaaaac tcgctctgta cggcttaggc
     3361 tccattccag atgaaaggct ctatcggatg tttgtgaata aaaaagtaac aatgttgaga
     3421 ccaaaggaag atgagaactc atggtttaac ttatttgtga ttcatcagaa caggagtaag
     3481 catggaaaca ccaacttcat tccagagcag tttttggatg acttcatcga cctcgttatc
     3541 tggggccatg aacatgagtg taaaattggc ccaatcaaaa atgagcagca gctcttctat
     3601 gtgtctcagc ccggaagctc agtggtgacg tccctttccc ctggagaagc tgtgaagaaa
     3661 catgtaggct tgctgcgcat taaagggaga aagatgaaca tgcagaagct gcctctccgc
     3721 accgtgcggc ggttcttcat agaagacgtg gttctggcta accacccaaa cctgttcaac
     3781 cctgacaatc ctaaggtgac ccaggccatc cagagcttct gtctggagaa gattgaagaa
     3841 atgcttgaca gtgctgagcg ggaacgactg gggaatcctc agcagccggg gaagcctctt
     3901 atccgactac gggtggacta tagtggaggc tttgaacctt tcaacgttct tcgttttagc
     3961 cagaagtttg tggatcgagt cgctaaccca aaagatgtca tccacttttt caggcacagg
     4021 gaacaaaagg gaaaaacagg agaagagatc aactttggga tgctcatcac aaaacctgct
     4081 tcagaaggag caacgcttag agtagaagac ctcgtaaagc agtatttcca aactgcagag
     4141 aagaatgttc agctctcgct gctgacagaa agagggatgg gcgaagcagt tcaagagttt
     4201 gtggacaagg aagagaaaga tgccatcgag gaattagtga agtaccagct ggagaaaaca
     4261 cagcggtttc ttaaggagcg ccatattgat gctctggaag acaagattga tgaggaggtc
     4321 cgacgtttcc gagaaagcag acagagaaat accaacgaag aagatgatga agttcgagag
     4381 gccatgagca gagcccgggc cctcagatca cagtcagaga cctccacctc agcctttagt
     4441 gctgaggacc tgagctttga tacatcggag cagacagcaa atgactctga tgacagcctg
     4501 tcagcagtgc cgagcagagg ccgaggccga ggccgagggc gaagaggagc cagagggcag
     4561 agctcggcac ctagaggagg ctctcagaga ggccgagaca ctgggctgga gatcactact
     4621 cgaggcagga gctcaaaggc cacctcatca acatctagaa acatgtccat tatagacgct
     4681 ttcagatcta cccgacaaca gccttctaga aatgtagccc ctaagaatta ctcagagacc
     4741 attgaggtgg atgactctga cgaagatgac atttttccta ccaattccag ggctgatcaa
     4801 aggtggtcgg gcacaacatc tagcaaacgg atgtcccaga gccagacagc caaaggggta
     4861 gactttgaat cagatgagga tgatgacgat gaccctttca tgagcagtag ttgcccaaga
     4921 agaaaccgaa gactcgagga ctacaaagac catgacggtg attataaaga tcatgacatc
     4981 gactacaagg atgacgatga caagtaggga tccgcccctc tccctccccc ccccctaacg
     5041 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     5101 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     5161 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     5221 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     5281 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     5341 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     5401 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     5461 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     5521 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     5581 gatgataagc ttgccacaac catggctgaa caagatggat tgcacgcagg ttctccggcc
     5641 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5701 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5761 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5821 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5881 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5941 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     6001 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     6061 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     6121 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     6181 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     6241 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     6301 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     6361 atcgccttct atcgccttct tgacgagttc ttctgaacgc gtctggaaca atcaacctct
     6421 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     6481 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     6541 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     6601 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     6661 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     6721 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     6781 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     6841 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     6901 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     6961 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt ctgcagtcga
     7021 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     7081 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     7141 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagagg
     7201 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     7261 cacacacaag gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat
     7321 ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag
     7381 gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac
     7441 ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc
     7501 cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt
     7561 tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc
     7621 agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat
     7681 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7741 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7801 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat
     7861 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc cttgacattg
     7921 ctagcgttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     8041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     8101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     8161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     8221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     8281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     8341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     8401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     8461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     8521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     8581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     8821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     9001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     9061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     9121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     9181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     9241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     9301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     9361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    10021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    10081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
    10141 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
    10201 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
    10261 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
    10321 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
    10381 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
    10441 t
//
