LOCUS       V014227                 8356 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014227
VERSION     V014227
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8356)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             234..414
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1076..1309
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /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             1686..1727
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    1798..1915
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2005..2208
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2297..2371
                     /label="COX8 presequence"
                     /note="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
     CDS             2405..2470
                     /label="3xFLAG"
                     /note="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
     promoter        2515..2726
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2739..3007
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3063..3728
                     /label="CopGFP"
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             3798..3851
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3852..4448
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4452..5039
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5113..5346
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5418..5552
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      5558..5693
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     primer_bind     complement(5731..5747)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5755..5771)
                     /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)."
     promoter        complement(5779..5809)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5824..5845)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6133..6721)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6895..7752)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7753..7857)
                     /label="AmpR promoter"
     primer_bind     8331..8347
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgt ccgtcctgac gccgctgctg ctgcggggct tgacaggctc
     2341 ggcccggcgg ctcccagtgc cgcgcgccaa gatccattcg ttgggggatc caccggtcgc
     2401 caccgactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2461 cgatgacaag tgagcggccg cgaaggatct gcgatcgctc cggtgcccgt cagtgggcag
     2521 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
     2581 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     2641 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     2701 gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc tctccttcac
     2761 gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt ctgccgcctc
     2821 ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag ctcaggtcga
     2881 gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg ctctccacgc
     2941 tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt ctgcgccgtt
     3001 acagatccaa gctgtgaccg gcgcctacgc tagacgccac catggagagc gacgagagcg
     3061 gcctgcccgc catggagatc gagtgccgca tcaccggcac cctgaacggc gtggagttcg
     3121 agctggtggg cggcggagag ggcaccccca agcagggccg catgaccaac aagatgaaga
     3181 gcaccaaagg cgccctgacc ttcagcccct acctgctgag ccacgtgatg ggctacggct
     3241 tctaccactt cggcacctac cccagcggct acgagaaccc cttcctgcac gccatcaaca
     3301 acggcggcta caccaacacc cgcatcgaga agtacgagga cggcggcgtg ctgcacgtga
     3361 gcttcagcta ccgctacgag gccggccgcg tgatcggcga cttcaaggtg gtgggcaccg
     3421 gcttccccga ggacagcgtg atcttcaccg acaagatcat ccgcagcaac gccaccgtgg
     3481 agcacctgca ccccatgggc gataacgtgc tggtgggcag cttcgcccgc accttcagcc
     3541 tgcgcgacgg cggctactac agcttcgtgg tggacagcca catgcacttc aagagcgcca
     3601 tccaccccag catcctgcag aacgggggcc ccatgttcgc cttccgccgc gtggaggagc
     3661 tgcacagcaa caccgagctg ggcatcgtgg agtaccagca cgccttcaag acccccatcg
     3721 ccttcgccag atcccgcgct cagtcgtcca attctgccgt ggacggcacc gccggacccg
     3781 gctccaccgg atctcgcgag ggcagaggaa gtcttctaac atgcggtgac gtggaggaga
     3841 atcccggccc tatgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     3901 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     3961 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4021 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4081 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4141 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4201 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4261 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4321 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4381 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     4441 ccggtgcctg aaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     4501 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     4561 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     4621 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     4681 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     4741 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     4801 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     4861 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     4921 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     4981 ttccgcgtct ccgccttcgc cctcagacga gtcggatctc cctttggccg cctccccgcc
     5041 tggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     5101 aaaagggggg actggaaggg ctaattcact cccaacgaaa ataagatctg ctttttgctt
     5161 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     5221 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     5281 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     5341 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     5401 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5461 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5521 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccag
     5581 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5641 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     5701 ttgcagagac ggcccaaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5761 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5821 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5881 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5941 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6001 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6061 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6121 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6181 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6241 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6301 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6361 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6421 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6481 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6541 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6601 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6661 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6721 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6781 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6841 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6901 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6961 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     7021 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     7081 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     7141 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     7201 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7261 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7321 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7381 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7441 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7501 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7561 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7621 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7681 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7741 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7801 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7861 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7921 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     8041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     8101 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     8161 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     8221 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     8281 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     8341 ggccagtgcc aagctg
//
