LOCUS       V023852                 9974 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023852
VERSION     V023852
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"
     promoter        2337..3519
                     /note="strong constitutive promoter for human elongation
                     factor EF-1α"
                     /label="EF-1α promoter"
     CDS             3551..3568
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             3569..4633
                     /label="CX3CR1(NM_001171172)"
                     /note="CX3CR1(NM_001171172)"
                     /gene="CX3CR1"
     CDS             4640..4705
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4724..5310
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5330..5929
                     /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    5943..6531
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6738..7371
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7499..7515)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7523..7539
                     /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(7547..7577)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7592..7613
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7901..8489)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8660..9520)
                     /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(9521..9625)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9673..9807
                     /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 cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtacgaatt
     3541 cgccaccatg caccatcacc atcaccatat ggatcagttc cctgaatcag tgacagaaaa
     3601 ctttgagtac gatgatttgg ctgaggcctg ttatattggg gacatcgtgg tctttgggac
     3661 tgtgttcctg tccatattct actccgtcat ctttgccatt ggcctggtgg gaaatttgtt
     3721 ggtagtgttt gccctcacca acagcaagaa gcccaagagt gtcaccgaca tttacctcct
     3781 gaacctggcc ttgtctgatc tgctgtttgt agccactttg cccttctgga ctcactattt
     3841 gataaatgaa aagggcctcc acaatgccat gtgcaaattc actaccgcct tcttcttcat
     3901 cggctttttt ggaagcatat tcttcatcac cgtcatcagc attgataggt acctggccat
     3961 cgtcctggcc gccaactcca tgaacaaccg gaccgtgcag catggcgtca ccatcagcct
     4021 aggcgtctgg gcagcagcca ttttggtggc agcaccccag ttcatgttca caaagcagaa
     4081 agaaaatgaa tgccttggtg actaccccga ggtcctccag gaaatctggc ccgtgctccg
     4141 caatgtggaa acaaattttc ttggcttcct actccccctg ctcattatga gttattgcta
     4201 cttcagaatc atccagacgc tgttttcctg caagaaccac aagaaagcca aagccattaa
     4261 actgatcctt ctggtggtca tcgtgttttt cctcttctgg acaccctaca acgttatgat
     4321 tttcctggag acgcttaagc tctatgactt ctttcccagt tgtgacatga ggaaggatct
     4381 gaggctggcc ctcagtgtga ctgagacggt tgcatttagc cattgttgcc tgaatcctct
     4441 catctatgca tttgctgggg agaagttcag aagatacctt taccacctgt atgggaaatg
     4501 cctggctgtc ctgtgtgggc gctcagtcca cgttgatttc tcctcatctg aatcacaaag
     4561 gagcaggcat ggaagtgttc tgagcagcaa ttttacttac cacacgagtg atggagatgc
     4621 attgctcctt ctctctagag actacaaaga ccatgacggt gattataaag atcatgacat
     4681 cgactacaag gatgacgatg acaagtagcg gccgcggatc ccgcccctct ccctcccccc
     4741 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     4801 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     4861 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     4921 atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga
     4981 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     5041 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     5101 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     5161 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     5221 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     5281 gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca tgaccgagta
     5341 caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc
     5401 cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc gccacatcga
     5461 gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt
     5521 gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc
     5581 gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc
     5641 cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt
     5701 cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt
     5761 gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc
     5821 gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt
     5881 gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga cgcgtctgga
     5941 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     6001 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     6061 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     6121 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     6181 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     6241 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     6301 tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc
     6361 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     6421 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     6481 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaatta
     6541 attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
     6601 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
     6661 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
     6721 aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
     6781 tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca caccagggcc
     6841 aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagccaga
     6901 taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg tgagcctgca
     6961 tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc gcctagcatt
     7021 tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat atcgagcttg
     7081 ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg ggactgggga
     7141 gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact gggtctctct
     7201 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
     7261 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
     7321 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtagtagtt
     7381 catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca gagagtgaga
     7441 ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     7501 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7561 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7681 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7741 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7801 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7861 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7921 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7981 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     8041 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     8101 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8161 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8221 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8281 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8341 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8401 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8461 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8521 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8581 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8641 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8701 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8761 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8821 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8881 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8941 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     9001 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     9061 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     9121 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9181 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9241 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9301 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9361 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9421 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9481 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9541 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9601 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     9661 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     9721 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9781 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     9841 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9901 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9961 acaaacttag tagt
//
