LOCUS       V015231                 9054 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V015231
VERSION     V015231
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9054)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        131..334
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             408..431
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             432..1322
                     /label="HaloTag(R)"
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
     CDS             1350..2066
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    2137..2254
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2303..2813
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2835..3431
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             3562..3795
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3867..4001
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4028..4163
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(4184..4202)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4212..4228)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      4370..4825
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4851..4955
                     /label="AmpR promoter"
     CDS             4956..5813
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5987..6575
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6863..6884
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6899..6929
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6937..6953
                     /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     6961..6977
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6998..7016
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7044..7270
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    7498..7623
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8116..8349
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8534..8578
                     /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             8727..8768
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      121 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      181 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      241 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      301 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      361 gcatggacta caaagaccat gacggtgatt ataaagatca tgacatcgat tacaaggatg
      421 acgatgacaa gatggcagaa atcggtactg gctttccatt cgacccccat tatgtggaag
      481 tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt
      541 tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac
      601 cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc
      661 tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc
      721 tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca
      781 agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga
      841 cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg
      901 tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg
      961 tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg
     1021 ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga
     1081 acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc
     1141 tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca
     1201 aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag
     1261 acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg
     1321 gcggcggagg gagtcaaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     1381 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1441 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1501 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1561 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1621 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1681 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1741 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1801 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1861 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1921 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1981 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2041 ctctcggcat ggacgagctg tacaagtccg gactcagatc tcgagctcaa gcttcgaatt
     2101 ctcgacctcg agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
     2161 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
     2221 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
     2281 tccactttgg ccgcggctcg agggggttgg ggttgcgcct tttccaaggc agccctgggt
     2341 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     2401 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     2461 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     2521 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     2581 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     2641 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     2701 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     2761 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     2821 caccggagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     2881 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     2941 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     3001 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     3061 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     3121 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     3181 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     3241 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     3301 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     3361 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     3421 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     3481 cccatgcatc ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact
     3541 ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc
     3601 tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     3661 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     3721 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     3781 ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta taacttgcaa
     3841 agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat ggttacaaat
     3901 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3961 gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc cgcccctaac
     4021 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4081 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4141 gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag tgagtcgtat
     4201 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4261 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4321 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4381 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4441 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4501 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4561 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4621 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4681 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4741 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4801 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4861 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4921 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4981 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5041 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5101 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5161 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5221 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5281 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5341 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5401 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5461 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5521 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5581 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5641 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5701 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5761 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5821 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6121 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6661 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6721 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6781 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     6841 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     6901 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     6961 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
     7021 aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt cttgcaacat
     7081 ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg catgccgatt
     7141 ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg gtctgacatg
     7201 gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt gcctagctcg
     7261 atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact
     7321 agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc
     7381 ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa
     7441 aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc
     7501 tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg
     7561 gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc
     7621 gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg
     7681 ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc
     7741 gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta
     7801 caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc
     7861 ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata
     7921 gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc
     7981 tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa
     8041 aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa
     8101 aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat
     8161 gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca
     8221 gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt
     8281 ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca
     8341 acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg
     8401 gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga
     8461 gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca
     8521 aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg
     8581 gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg
     8641 aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca
     8701 gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc
     8761 cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa
     8821 cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct tcaccgaggg
     8881 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     8941 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     9001 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atca
//
