LOCUS       V003619                 8600 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003619
VERSION     V003619
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8600)
  AUTHORS   Cordero Llana O.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-APR-2014) Physiology Anatomy and Genetics, University
            of Oxford, South Parks Rd., Oxford, England OX1 3QX, United Kingdom
REFERENCE   2  (bases 1 to 8600)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8600)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted
            (12-APR-2014) Physiology Anatomy and Genetics, University of Oxford,
            South Parks Rd., Oxford, England OX1 3QX, United Kingdom"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    52..640
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     misc_feature    710..725
                     /label="cPPPT5'"
                     /note="cPPPT5'"
     LTR             727..960
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1032..1166
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1172..1307
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1340..1358)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1368..1384)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1529..1984
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2013..2117
                     /label="AmpR promoter"
     CDS             2118..2975
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      3149..3737
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4025..4046
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4061..4091
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4099..4115
                     /label="lac operator"
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4123..4139
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4160..4178
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4206..4433
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    4434..4614
                     /label="HIV-LTR"
                     /note="HIV-LTR"
     LTR             4434..4614
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4658..4783
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5276..5509
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5694..5738
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             5887..5928
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    5999..6116
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6233..6640
                     /label="SFFV promoter"
                     /note="spleen focus-forming virus long terminal repeat
                     (LTR) promoter"
     CDS             6662..7378
                     /label="EGFP"
                     /note="enhanced GFP"
     enhancer        7458..7761
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7762..7965
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             8060..8596
                     /gene="MANF"
                     /label="Mesencephalic astrocyte-derived neurotrophic
                     factor"
                     /note="Mesencephalic astrocyte-derived neurotrophic factor
                     from Homo sapiens. Accession#: P55145"
ORIGIN
        1 ctagtggatc atctttgtcg actagggata acagggtaat gctagctcga caatcaacct
       61 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
      121 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
      181 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
      241 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
      301 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
      361 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
      421 gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt
      481 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
      541 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
      601 cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg agctcggtac
      661 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
      721 ggggactgga agggctaatt cactcccaac gaagacaaga tctgcttttt gcttgtactg
      781 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
      841 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
      901 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
      961 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     1021 agagtgagag gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     1081 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     1141 tcaatgtatc ttatcatgtc tggctctagc tatcccgccc ctaactccgc ccagttccgc
     1201 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1261 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcgt
     1321 cgagacgtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt
     1381 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     1441 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1501 ttgcgcagcc tgaatggcga atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg
     1561 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     1621 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     1681 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     1741 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1801 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1861 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1921 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     1981 atttcccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2041 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2101 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2161 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2221 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2281 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2341 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2401 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2461 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2521 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     2581 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     2641 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     2701 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     2761 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     2821 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     2881 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     2941 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     3001 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     3061 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3121 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3181 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3241 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3301 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3361 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3421 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3481 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3541 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3601 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3661 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3721 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3781 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3841 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     3901 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     3961 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4021 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     4081 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     4141 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctgcaag
     4201 cttaatgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
     4261 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
     4321 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
     4381 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
     4441 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     4501 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     4561 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     4621 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
     4681 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
     4741 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
     4801 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
     4861 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
     4921 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
     4981 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
     5041 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
     5101 taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg
     5161 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     5221 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     5281 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     5341 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     5401 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     5461 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     5521 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     5581 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca
     5641 attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg
     5701 aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa
     5761 attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa
     5821 tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt
     5881 ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag
     5941 gtggagagag agacagagac agatccattc gattagtgaa cggatctcga cggttaactt
     6001 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
     6061 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttatcg
     6121 ataagcttga tatcgaattc ctgcagcccc gataaaataa aagattttat ttagtctcca
     6181 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctg cagtaacgcc
     6241 attttgcaag gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg
     6301 tacatgaaaa tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc
     6361 ggcccggggc caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca
     6421 gatggtcccc agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca
     6481 ggctccccca aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct
     6541 tctcgcttct gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc
     6601 actcggcgcg ccagtcctcc gacagactga gtcgcccggg ggggatccac cggtcgccac
     6661 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6721 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     6781 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     6841 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     6901 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     6961 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     7021 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     7081 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     7141 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     7201 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     7261 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     7321 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     7381 aagcggccgc gactctagag tcgacctgca ggcatgcaag cttgatatca agcttatcga
     7441 taagcttggg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     7501 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     7561 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     7621 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     7681 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     7741 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     7801 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     7861 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     7921 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt
     7981 cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgactctag
     8041 atgcatgctc gaggccacca tgtgggccac gcaggggctg gcggtggcgc tggctctgag
     8101 cgtgctgccg ggcagccggg cgctgcggcc gggcgactgc gaagtttgta tttcttatct
     8161 gggaagattt taccaggacc tcaaagacag agatgtcaca ttctcaccag ccactattga
     8221 aaacgaactt ataaagttct gccgggaagc aagaggcaaa gagaatcggt tgtgctacta
     8281 tatcggggcc acagatgatg cagccaccaa aatcatcaat gaggtatcaa agcctctggc
     8341 ccaccacatc cctgtggaga agatctgtga gaagcttaag aagaaggaca gccagatatg
     8401 tgagcttaag tatgacaagc agatcgacct gagcacagtg gacctgaaga agctccgagt
     8461 taaagagctg aagaagattc tggatgactg gggggagaca tgcaaaggct gtgcagaaaa
     8521 gtctgactac atccggaaga taaatgaact gatgcctaaa tatgccccca aggcagccag
     8581 tgcacggacc gatttgtaga
//
