LOCUS       .                       9958 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        6..233
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    458..583
                     /label="HIV-1 Ψ"
                     /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
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    1804..1920
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2010..2213
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2302..3018
                     /label="EYFP"
                     /note="enhanced YFP"
     CDS             3070..3087
                     /label="6xHis"
                     /note="6xHis affinity tag"
     promoter        3130..3341
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     LTR             3354..3622
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3681..5330
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             5400..5453
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5454..6050
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    6054..6641
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6715..6948
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7020..7154
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7160..7295
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(7381..7411)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7426..7447)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7735..8323)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8497..9354)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(9355..9459)
                     /label="AmpR promoter"
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 ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaat tcaaaatttt
     1921 atcgatacta gtattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1981 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     2041 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     2101 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     2161 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     2221 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagattct
     2281 agagctagcg aattcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     2341 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2401 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2461 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     2521 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2581 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2641 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2701 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2761 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2821 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2881 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     2941 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     3001 atggacgagc tgtacaaggg cggtggagga tcaggtggag gcggatccgc tcgaattaaa
     3061 tcactcgagc accatcacca tcaccattag cggccgcaag gatctgcgat cgctccggtg
     3121 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
     3181 gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
     3241 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
     3301 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc
     3361 gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc
     3421 gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt
     3481 taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca
     3541 gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt
     3601 ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac gccaccatgg
     3661 agagcgacga gagcggcctg atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
     3721 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
     3781 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
     3841 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
     3901 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     3961 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     4021 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     4081 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     4141 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     4201 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     4261 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     4321 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     4381 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     4441 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     4501 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     4561 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     4621 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     4681 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacacccg
     4741 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     4801 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     4861 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     4921 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     4981 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     5041 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     5101 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     5161 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     5221 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     5281 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg agatcccgcg
     5341 ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc ggatctcgcg
     5401 agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc cctatgaccg
     5461 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5521 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5581 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5641 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5701 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5761 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5821 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5881 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5941 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     6001 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac
     6061 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     6121 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     6181 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     6241 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     6301 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     6361 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     6421 ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg
     6481 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     6541 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc
     6601 gccctcagac gagtcggatc tccctttggc cgcctccccg cctggtacct ttaagaccaa
     6661 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6721 ggctaattca ctcccaacga aaataagatc tgctttttgc ttgtactggg tctctctggt
     6781 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     6841 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     6901 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     6961 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
     7021 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     7081 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     7141 atcatgtctg gctctagcta tcccgcccct aactccgccc agttccgccc attctccgcc
     7201 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7261 attccagaag tagtgaggag gcttttttgg aggcctagac ttttgcagag acggcccaaa
     7321 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     7381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     7441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7501 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7561 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7621 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7681 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7741 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7801 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7861 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7921 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7981 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     8041 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     8101 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     8161 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     8221 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     8281 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     8341 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     8401 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     8461 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     8521 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     8581 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     8641 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     8701 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     8761 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     8821 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     8881 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     8941 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     9001 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     9061 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     9121 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     9181 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     9241 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     9301 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     9361 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     9421 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     9481 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     9541 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     9601 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     9661 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     9721 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     9781 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     9841 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     9901 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctg
//
