LOCUS       V033896                 9382 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033896
VERSION     V033896
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..4078
                     /label="Tgfb3(NM_009368)"
                     /note="Tgfb3(NM_009368)"
                     /gene="Tgfb3"
     CDS             4085..4150
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        4170..4669
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4690..5289
                     /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    5303..5891
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6098..6731
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6860..6876)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6884..6900
                     /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(6908..6938)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6953..6974
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7262..7850)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8021..8881)
                     /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(8882..8986)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9034..9168
                     /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 ccatgaagat gcacttgcaa agggctctgg tagtcctggc
     2881 cctgctgaac ttggccacaa tcagcctctc tctgtccact tgcaccacgt tggacttcgg
     2941 ccacatcaag aagaagaggg tggaagccat taggggacag atcttgagca agctcaggct
     3001 caccagcccc cctgagccat cggtgatgac ccacgtcccc tatcaggtcc tggcacttta
     3061 caacagcacc cgggagttgc tggaagagat gcacggggag agggaggaag gctgcactca
     3121 ggagacctcg gagtctgagt actatgccaa agagatccat aaattcgaca tgatccaggg
     3181 actggcggag cacaatgaac tggccgtctg ccccaaagga attacctcta aggtttttcg
     3241 tttcaatgtg tcctcagtgg agaaaaatgg aaccaatctg ttccgggcag agttccgggt
     3301 cttgcgggtg cccaacccca gctccaagcg cacagagcag agaattgagc tcttccagat
     3361 acttcgaccg gatgagcaca tagccaagca gcgctacata ggtggcaaga atctgcccac
     3421 aaggggcacc gctgaatggc tgtctttcga tgtcactgac actgtgcgcg agtggctgtt
     3481 gaggagagag tccaacttgg gtctggaaat cagcatccac tgtccatgtc acacctttca
     3541 gcccaatgga gacatactgg aaaatgttca tgaggtgatg gaaatcaaat tcaaaggagt
     3601 ggacaatgaa gatgaccatg gccgtggaga cctggggcgt ctcaagaagc aaaaggatca
     3661 ccacaaccca cacctgatcc tcatgatgat ccccccacac cgactggaca gcccaggcca
     3721 gggcagtcag aggaagaaga gggccctgga caccaattac tgcttccgca acctggagga
     3781 gaactgctgt gtacgccccc tttatattga cttccggcag gatctaggct ggaaatgggt
     3841 ccacgaacct aagggttact atgccaactt ctgctcaggc ccttgcccat acctccgcag
     3901 cgcagacaca acccatagca cggtgcttgg actatacaac accctgaacc cagaggcgtc
     3961 tgcctcgcca tgctgcgtcc cccaggacct ggagcccctg accatcttgt actatgtggg
     4021 cagaaccccc aaggtggagc agctgtccaa catggtggtg aagtcgtgta agtgcagcgg
     4081 atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     4141 cgatgacaag tagtctagat aattctaccg ggtaggggag gcgcttttcc caaggcagtc
     4201 tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg gcctctggcc
     4261 tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg gtggcccctt
     4321 cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc agctcgcgtc
     4381 gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga tggacagcac
     4441 cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc tttgctcctt
     4501 cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca ggggcgggct
     4561 caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca cgcttcaaaa
     4621 gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga cctgcagccc
     4681 aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     4741 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     4801 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     4861 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     4921 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     4981 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     5041 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     5101 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     5161 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     5221 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     5281 ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac
     5341 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     5401 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     5461 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     5521 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     5581 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     5641 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     5701 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     5761 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     5821 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     5881 cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca
     5941 agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag
     6001 gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct
     6061 gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa
     6121 cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt ccctgattag
     6181 cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag
     6241 ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg
     6301 ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg
     6361 tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag
     6421 aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt
     6481 ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt
     6541 tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     6601 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     6661 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     6721 aatctctagc agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
     6781 atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg acctctagct
     6841 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6901 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6961 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8161 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8881 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9001 aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc ttataatggt
     9061 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9121 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact
     9181 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac
     9241 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat
     9301 tgtatttgtt aaatatgtac tacaaactta gtagttttta aagaaattgt atttgttaaa
     9361 tatgtactac aaacttagta gt
//
