LOCUS       V019414                 9867 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019414
VERSION     V019414
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             2827..4593
                     /label="IL4I1(NM_001258017)"
                     /note="IL4I1(NM_001258017)"
                     /gene="IL4I1"
     CDS             4600..4623
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4655..5154
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5175..5774
                     /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    5788..6376
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6583..7216
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7345..7361)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7369..7385
                     /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(7393..7423)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7438..7459
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7747..8335)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8506..9366)
                     /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(9367..9471)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9519..9653
                     /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 gccaccatgc ccaacgatga cttctgtcct gggctaacca taaaggccat gggtgctgag
     2881 agagcccccc agaggcagcc atgcaccctg cacctcctcg tcctcgtccc catcctcctc
     2941 agcctggtgg cctcccagga ctggaaggct gaacgcagcc aagacccctt cgagaaatgc
     3001 atgcaggatc ctgactatga gcagctgctc aaggtggtga cctgggggct caatcggacc
     3061 ctgaagcccc agagggtgat tgtggttggc gctggtgtgg ccgggctggt ggccgccaag
     3121 gtgctcagcg atgctggaca caaggtcacc atcctggagg cagataacag gatcgggggc
     3181 cgcatcttca cctaccggga ccagaacacg ggctggattg gggagctggg agccatgcgc
     3241 atgcccagct ctcacaggat cctccacaag ctctgccagg gcctggggct caacctgacc
     3301 aagttcaccc agtacgacaa gaacacgtgg acggaggtgc acgaagtgaa gctgcgcaac
     3361 tatgtggtgg agaaggtgcc cgagaagctg ggctacgcct tgcgtcccca ggaaaagggc
     3421 cactcgcccg aagacatcta ccagatggct ctcaaccagg ccctcaaaga cctcaaggca
     3481 ctgggctgca gaaaggcgat gaagaagttt gaaaggcaca cgctcttgga atatcttctc
     3541 ggggagggga acctgagccg gccggccgtg cagcttctgg gagacgtgat gtccgaggat
     3601 ggcttcttct atctcagctt cgccgaggcc ctccgggccc acagctgcct cagcgacaga
     3661 ctccagtaca gccgcatcgt gggtggctgg gacctgctgc cgcgcgcgct gctgagctcg
     3721 ctgtccgggc ttgtgctgtt gaacgcgccc gtggtggcga tgacccaggg accgcacgat
     3781 gtgcacgtgc agatcgagac ctctcccccg gcgcggaatc tgaaggtgct gaaggccgac
     3841 gtggtgctgc tgacggcgag cggaccggcg gtgaagcgca tcaccttctc gccgccgctg
     3901 ccccgccaca tgcaggaggc gctgcggagg ctgcactacg tgccggccac caaggtgttc
     3961 ctaagcttcc gcaggccctt ctggcgcgag gagcacattg aaggcggcca ctcaaacacc
     4021 gatcgcccgt cgcgcatgat tttctacccg ccgccgcgcg agggcgcgct gctgctggcc
     4081 tcgtacacgt ggtcggacgc ggcggcagcg ttcgccggct tgagccggga agaggcgttg
     4141 cgcttggcgc tcgacgacgt ggcggcattg cacgggcctg tcgtgcgcca gctctgggac
     4201 ggcaccggcg tcgtcaagcg ttgggcggag gaccagcaca gccagggtgg ctttgtggta
     4261 cagccgccgg cgctctggca aaccgaaaag gatgactgga cggtccctta tggccgcatc
     4321 tactttgccg gcgagcacac cgcctacccg cacggctggg tggagacggc ggtcaagtcg
     4381 gcgctgcgcg ccgccatcaa gatcaacagc cggaaggggc ctgcatcgga cacggccagc
     4441 cccgaggggc acgcatctga catggagggg caggggcatg tgcatggggt ggccagcagc
     4501 ccctcgcatg acctggcaaa ggaagaaggc agccaccctc cagtccaagg ccagttatct
     4561 ctccaaaaca cgacccacac gaggacctcg catgaattcg attacaagga tgacgacgat
     4621 aagtgaggat cccgcgactc tagataattc taccgggtag gggaggcgct tttcccaagg
     4681 cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc
     4741 tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc
     4801 cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc
     4861 gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac
     4921 agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc
     4981 tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc
     5041 gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt
     5101 caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc
     5161 agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5221 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5281 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5341 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5401 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5461 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5521 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5581 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5641 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5701 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5761 agcccggtgc ctgaccgcgt ctggaacaat caacctctgg attacaaaat ttgtgaaaga
     5821 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     5881 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     5941 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     6001 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     6061 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     6121 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     6181 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     6241 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     6301 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     6361 tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa
     6421 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     6481 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     6541 cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg ctaattcact
     6601 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     6661 attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt ggatggtgct
     6721 acaagctagt accagttgag ccagataagg tagaagaggc caataaagga gagaacacca
     6781 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtgttagagt
     6841 ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat ccggagtact
     6901 tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac tttccaggga
     6961 ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata taagcagctg
     7021 ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     7081 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     7141 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     7201 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     7261 aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgttttac cgtcgacctc
     7321 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     7381 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7441 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7501 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7561 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7621 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7681 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7741 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7801 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7861 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7921 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7981 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     8041 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     8101 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     8161 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     8221 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     8281 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     8341 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     8401 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8461 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8641 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9361 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9421 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9481 ccgaaaagtg ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata
     9541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     9601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga
     9661 taactcaagc taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca
     9721 attacctgtg gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa
     9781 gaaattgtat ttgttaaata tgtactacaa acttagtagt ttttaaagaa attgtatttg
     9841 ttaaatatgt actacaaact tagtagt
//
