LOCUS       V037500                 9961 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037500
VERSION     V037500
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..3559
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             3629..4729
                     /label="Ccr6(NM_009835)"
                     /note="Ccr6(NM_009835)"
                     /gene="Ccr6"
     promoter        4749..5248
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5269..5868
                     /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    5882..6470
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6677..7310
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7439..7455)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7463..7479
                     /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(7487..7517)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7532..7553
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7841..8429)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8600..9460)
                     /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(9461..9565)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9613..9747
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt
     2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg
     3601 gggaagcgga ggcggaggga gtggatccat gaattccaca gagtcctact ttggaacgga
     3661 tgattatgac aacacagagt attattctat tcctccagac catgggccat gctccctaga
     3721 agaggtcaga aacttcacca aggtatttgt gccaattgcc tactccttaa tatgtgtctt
     3781 tggcctcctg ggcaacatta tggtggtgat gacctttgcc ttctacaaga aagccagatc
     3841 catgactgac gtctacctgt tgaacatggc catcacagac atactctttg tcctcaccct
     3901 accgttctgg gcagttactc atgccaccaa cacttgggtt ttcagcgatg cactgtgtaa
     3961 actgatgaaa ggcacatatg cggtcaactt taactgtggg atgctgctcc tggcctgtat
     4021 cagcatggac cggtacattg ccatcgtcca ggcaaccaaa tctttccggg tacgctccag
     4081 aacactgacg cacagtaagg tcatctgtgt ggcagtgtgg ttcatctcca tcatcatctc
     4141 aagccctaca tttatcttca acaagaaata cgagctgcag gatcgtgatg tctgtgagcc
     4201 acggtacagg tctgtctcag agcccatcac gtggaagctg ctgggtatgg gactggagct
     4261 gttctttggg ttcttcaccc ctttgctgtt tatggtgttc tgctatctgt tcattatcaa
     4321 gaccttggtg caggcccaga actccaagag gcacagagca atccgagtcg tgatcgctgt
     4381 ggttctcgtg ttcctggctt gtcagatccc tcacaacatg gtcctcctcg tgactgcggt
     4441 caacacgggc aaagtgggcc ggagctgcag caccgagaaa gtcctcgcct acaccaggaa
     4501 cgtggccgag gtcctggctt tcctgcattg ctgcctcaac cccgtgttgt atgcgtttat
     4561 tggacagaaa ttcagaaact acttcatgaa gatcatgaag gatgtgtggt gtatgagaag
     4621 gaagaataag atgcctggct tcctctgtgc ccgggtttac tcggaaagct acatctccag
     4681 gcagaccagt gagaccgtcg aaaatgataa tgcatcgtcc tttaccatgt aatctagata
     4741 attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc
     4801 cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac
     4861 cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc
     4921 cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga
     4981 agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt
     5041 aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct
     5101 gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg
     5161 aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt
     5221 ctcctcttcc tcatctccgg gcctttcgac ctgcagccca agcttaccat gaccgagtac
     5281 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     5341 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     5401 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5461 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5521 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5581 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5641 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5701 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5761 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5821 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacc gcgtctggaa
     5881 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     5941 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6001 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6061 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6121 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6181 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6241 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6301 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     6361 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     6421 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattaa
     6481 ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6541 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     6601 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     6661 aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     6721 ctgtggatct accacacaca aggctacttc cctgattagc agaactacac accagggcca
     6781 ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagccagat
     6841 aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt gagcctgcat
     6901 gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg cctagcattt
     6961 catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata tcgagcttgc
     7021 tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg gactggggag
     7081 tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg ggtctctctg
     7141 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
     7201 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
     7261 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc
     7321 atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag
     7381 gccttgacat tgctagcgtt ttaccgtcga cctctagcta gagcttggcg taatcatggt
     7441 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7501 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7561 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7621 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7681 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7741 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7801 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7861 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7921 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7981 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     8041 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8101 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8161 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8221 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8281 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8341 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8401 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8461 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8521 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8581 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8641 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8701 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8761 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8821 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8881 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8941 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     9001 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     9061 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9121 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9181 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9241 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9301 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9361 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9421 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9481 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9541 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     9601 gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     9661 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     9721 atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca aaatcatccc
     9781 aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca tttactctaa
     9841 acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta aatatgtact
     9901 acaaacttag tagtttttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9961 t
//
