LOCUS       62056_14215        8037 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8037)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8037
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        256..287
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             333..1049
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    1120..1237
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        1286..1796
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             1818..2414
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     LTR             2545..2778
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2850..2984
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3011..3146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3167..3185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3195..3211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3353..3808
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3834..3938
                     /label=AmpR promoter
     CDS             3939..4796
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4970..5558
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5846..5867
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5882..5912
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5920..5936
                     /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     5944..5960
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5981..5999
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6027..6253
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6254..6434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6481..6606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7099..7332
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7517..7561
                     /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             7710..7751
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
ORIGIN
        1 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
       61 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtaa ggatgtccat
      121 attaggacat ctaggatgtc catattagga catctaggat gtccatatta ggacatctag
      181 gatgtccata ttaggacatc taggatgtcc atattaggac atctagatct ggcctcggcg
      241 gccaagctta gacactagag ggtatataat ggaagctcga cttccagctt ggcaatccgg
      301 tactgttggt aaagccacca ccggtcgcca ccatggtgag caagggcgag gagctgttca
      361 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      421 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      481 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      541 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      601 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      661 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      721 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      781 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      841 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      901 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
      961 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1021 tcactctcgg catggacgag ctgtacaagt ccggactcag atctcgagct caagcttcga
     1081 attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     1141 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     1201 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     1261 agatccactt tggccgcggc tcgagggggt tggggttgcg ccttttccaa ggcagccctg
     1321 ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc
     1381 ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc gccgctaccc
     1441 ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt tccttgcggt
     1501 tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc ctcgcagacg
     1561 gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct
     1621 gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg
     1681 gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg
     1741 agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct ccccaggggg
     1801 atccaccgga gcttaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     1861 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     1921 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     1981 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     2041 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     2101 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
     2161 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     2221 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
     2281 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     2341 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     2401 gcaagcccgg tgcctgacgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     2461 gaccccatgc atcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2521 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     2581 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2641 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2701 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     2761 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     2821 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     2881 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2941 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     3001 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3061 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3121 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     3181 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     3241 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     3301 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     3361 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3421 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3481 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3541 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3601 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3661 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3721 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3781 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     3841 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3901 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3961 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4021 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4081 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4141 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4201 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4261 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4321 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4381 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4441 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4501 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4561 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4621 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4681 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4741 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4801 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4861 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4921 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4981 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5041 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5101 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5161 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5221 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5281 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5341 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5401 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5461 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5521 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5581 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5641 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     5701 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     5761 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     5821 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5881 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5941 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
     6001 aacaaaagct ggagctgcaa gcttaatgta gtcttatgca atactcttgt agtcttgcaa
     6061 catggtaacg atgagttagc aacatgcctt acaaggagag aaaaagcacc gtgcatgccg
     6121 attggtggaa gtaaggtggt acgatcgtgc cttattagga aggcaacaga cgggtctgac
     6181 atggattgga cgaaccactg aattgccgca ttgcagagat attgtattta agtgcctagc
     6241 tcgatacata aacgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6301 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6361 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6421 gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag
     6481 ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga
     6541 ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag
     6601 agcgtcagta ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca
     6661 gggggaaaga aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga
     6721 ttcgcagtta atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag
     6781 ctacaaccat cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca
     6841 accctctatt gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag
     6901 atagaggaag agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag
     6961 acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt
     7021 aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga
     7081 aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac
     7141 tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt
     7201 gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac
     7261 agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga
     7321 tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc
     7381 ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat
     7441 ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc
     7501 gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt
     7561 gtggaattgg tttaacataa caaattggct gtggtatata aaattattca taatgatagt
     7621 aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag
     7681 gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag
     7741 gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt
     7801 gaacggatct cgacggtatc gatcacgaga ctagcctcga gcggccgccc ccttcaccga
     7861 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
     7921 aattggaatt aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa
     7981 gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatca
//
