LOCUS       .                       8970 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        7..507
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             522..917
                     /label="BSD"
                     /note="blasticidin S deaminase"
     LTR             992..1225
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1297..1431
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      1458..1593
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(1614..1632)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1642..1658)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      1800..2255
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2281..2385
                     /label="AmpR promoter"
     CDS             2386..3243
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      3417..4005
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    4293..4314
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4329..4359
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4367..4383
                     /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-β-D-thiogalactopyranoside (IPTG)."
     primer_bind     4391..4407
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        4428..4446
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4474..4700
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    4928..5053
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    5546..5779
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             5964..6008
                     /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             6157..6198
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     promoter        6306..6546
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        6568..6871
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6872..7075
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             7110..8759
                     /label="luciferase"
                     /note="firefly luciferase"
     misc_feature    8801..8918
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        8967..9477
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatccgccac catggccaag cctttgtctc
      541 aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg
      601 aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca
      661 atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg
      721 ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct
      781 tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca
      841 tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg
      901 gttatgtgtg ggagggctaa ggtaccttta agaccaatga cttacaaggc agctgtagat
      961 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     1021 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     1081 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1141 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1201 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     1261 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     1321 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1381 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     1441 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     1501 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     1561 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     1621 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1681 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1741 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     1801 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1861 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1921 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1981 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2041 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2101 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2161 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2221 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     2401 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     2461 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     2521 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     2581 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     2641 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     2701 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     2761 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     2821 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     2881 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     2941 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3001 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3061 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3121 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3181 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3241 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3301 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3361 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3541 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     3601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     3661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     3721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3781 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4021 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4081 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4141 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4201 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4261 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4321 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4381 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     4441 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     4501 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     4561 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     4621 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     4681 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     4741 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     4801 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     4861 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     4921 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     4981 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     5041 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     5101 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     5161 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     5221 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     5281 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     5341 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     5401 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     5461 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     5521 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     5581 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     5641 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     5701 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     5761 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     5821 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     5881 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     5941 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     6001 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     6061 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     6121 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     6181 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     6241 gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct
     6301 tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     6361 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     6421 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     6481 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     6541 aaggacgaaa caccggtcgg aattccgcgt tacataactt acggtaaatg gcccgcctgg
     6601 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6661 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6721 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6781 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6841 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     6901 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     6961 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7021 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctaatcg
     7081 tcgacgattc gtcagaatcg gtcgccacca tggaggacgc caagaacatc aagaagggcc
     7141 ccgccccctt ctaccccctg gaggacggca ccgccggcga gcagctgcac aaggccatga
     7201 agaggtacgc cctggtgccc ggcaccatcg ccttcaccga cgcccacatc gaggtgaaca
     7261 tcacctacgc cgagtacttc gagatgagcg tgaggctggc cgaggccatg aagaggtacg
     7321 gcctgaacac caaccacagg atcgtggtgt gcagcgagaa cagcctgcag ttcttcatgc
     7381 ccgtgctggg cgccctgttc atcggcgtgg ccgtggcccc cgccaacgac atctacaacg
     7441 agagggagct gctgaacagc atgaacatca gccagcccac cgtggtgttc gtgagcaaga
     7501 agggcctgca gaagatcctg aacgtgcaga agaagctgcc catcatccag aagatcatca
     7561 tcatggacag caagaccgac taccagggct tccagagcat gtacaccttc gtgaccagcc
     7621 acctgccccc cggcttcaac gagtacgact tcgtgcccga gagcttcgac agggacaaga
     7681 ccatcgccct gatcatgaac agcagcggca gcaccggcct gcccaagggc gtggccctgc
     7741 cccacaggac cgcctgcgtg aggttcagcc acgccaggga ccccatcttc ggcaaccaga
     7801 tcatccccga caccgccatc ctgagcgtgg tgcccttcca ccacggcttc ggcatgttca
     7861 ccaccctggg ctacctgatc tgcggcttca gggtggtgct gatgtacagg ttcgaggagg
     7921 agctgttcct gaggagcctg caggactaca agatccagag cgccctgctg gtgcccaccc
     7981 tgttcagctt cttcgccaag agcaccctga tcgacaagta cgacctgagc aacctgcacg
     8041 agatcgccag cggcggcgcc cccctgagca aggaggtggg cgaggccgtg gccaagaggt
     8101 tccacctgcc cggcatcagg cagggctacg gcctgaccga gaccaccagc gccatcctga
     8161 tcacccccga gggcgacgac aagcccggcg ccgtgggcaa ggtggtgccc ttcttcgagg
     8221 ccaaggtggt ggacctggac accggcaaga ccctgggcgt gaaccagagg ggcgagctgt
     8281 gcgtgagggg ccccatgatc atgagcggct acgtgaacaa ccccgaggcc accaacgccc
     8341 tgatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac gaggacgagc
     8401 acttcttcat cgtggacagg ctgaagagcc tgatcaagta caagggctac caggtggccc
     8461 ccgccgagct ggagagcatc ctgctgcagc accccaacat cttcgacgcc ggcgtggccg
     8521 gcctgcccga cgacgacgcc ggcgagctgc ccgccgccgt ggtggtgctg gagcacggca
     8581 agaccatgac cgagaaggag atcgtggact acgtggccag ccaggtgacc accgccaaga
     8641 agctgagggg cggcgtggtg ttcgtggacg aggtgcccaa gggcctgacc ggcaagctgg
     8701 acgccaggaa gatcagggag atcctgatca aggccaagaa gggcggcaag agcaagctgt
     8761 aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga
     8821 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta
     8881 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca
     8941 gagatccact ttggccgcgg ctcgaggggg
//
