LOCUS       .                       9621 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     polyA_signal    221..355
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      382..517
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(538..556)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(566..582)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      724..1179
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1205..1309
                     /label="AmpR promoter"
     CDS             1310..2167
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      2341..2929
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    3217..3238
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3253..3283
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    3291..3307
                     /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     3315..3331
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        3352..3370
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        3398..3624
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             3625..3805
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    3852..3977
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    4470..4703
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             4888..4932
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     promoter        5163..5373
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     CDS             5413..6009
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    6058..6175
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6230..6733
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    6754..6878
                     /label="attR1"
                     /note="recombination site for the Gateway® LR reaction"
     promoter        6903..6933
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6987..7643
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             7988..8290
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8334..8458)
                     /label="attR2"
                     /note="recombination site for the Gateway® LR reaction"
     CDS             8466..8786
                     /label="DmrB"
                     /note="F36V mutant of FK506-binding protein FKBP12"
     CDS             8793..8819
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8823..8849
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    8871..9459
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
ORIGIN
        1 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
       61 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
      121 gtcagtgtgg aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta
      181 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
      241 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      301 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
      361 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      421 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      481 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
      541 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      601 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      661 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      721 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      781 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      841 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      901 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      961 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1021 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1081 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1141 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1201 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1261 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1321 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1381 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1441 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1501 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1561 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1621 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1681 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1741 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1801 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1861 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1921 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1981 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2041 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2101 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2161 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2221 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2281 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2341 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2401 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2461 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2521 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2581 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2641 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2701 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2761 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2821 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2881 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2941 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3001 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3061 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3121 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3181 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3241 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3301 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     3361 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     3421 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     3481 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     3541 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     3601 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     3661 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3721 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3781 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     3841 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     3901 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     3961 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     4021 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     4081 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     4141 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     4201 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     4261 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     4321 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     4381 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     4441 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     4501 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     4561 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     4621 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     4681 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     4741 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     4801 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     4861 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     4921 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     4981 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     5041 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     5101 ggacccatgc attgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5161 cagcaggcag aagtatgcaa agcatgcgtc tcaattagtc agcaaccata gtcccgcccc
     5221 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5281 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5341 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttct agaggtacca
     5401 ccggagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5461 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5521 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5581 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgccgt ggcggtctgg
     5641 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5701 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5761 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5821 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     5881 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     5941 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6001 cccggtgcca gttaacgaat tctagatctt gagacaaatg gcagtattca tccacaattt
     6061 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     6121 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttcgggt
     6181 ttattacagg gacagcagag atccactttg gcgccggctc gaggcctgca ggttgcgcct
     6241 tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa
     6301 acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca gcgtcacccg
     6361 gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg
     6421 ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg cacgtctcac
     6481 tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac cgcgatgggc
     6541 tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg aaggggcggt
     6601 gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc
     6661 attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga
     6721 cctctctccc caggggctag catcgattga tcaacaagtt tgtacaaaaa agctgaacga
     6781 gaaacgtaaa atgatataaa tatcaatata ttaaattaga ttttgcataa aaaacagact
     6841 acataatact gtaaaacaca acatatccag tcactatggc ggccgcatta ggcaccccag
     6901 gctttacact ttatgcttcc ggctcgtata atgtgtggat tttgagttag gatccgtcga
     6961 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     7021 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     7081 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     7141 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     7201 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     7261 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     7321 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     7381 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     7441 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     7501 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     7561 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     7621 gcgatgagtg gcagggcggg gcgtaaagat ctggatccgg cttactaaaa gccagataac
     7681 agtatgcgta tttgcgcgct gatttttgcg gtataagaat atatactgat atgtataccc
     7741 gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt acagtgacag ttgacagcga
     7801 cagctatcag ttgctcaagg catatatgat gtcaatatct ccggtctggt aagcacaacc
     7861 atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga aagcggaaaa tcaggaaggg
     7921 atggctgagg tcgcccggtt tattgaaatg aacggctctt ttgctgacga gaacaggggc
     7981 tggtgaaatg cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt
     8041 ggatgtacag agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag
     8101 tgcacgtctg ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga
     8161 tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggga
     8221 agaagtggct gatctcagcc accgcgaaaa tgacatcaaa aacgccatta acctgatgtt
     8281 ctggggaata taaatgtcag gctcccttat acacagccag tctgcaggtc gaccatagtg
     8341 actggatatg ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta
     8401 atatattgat atttatatca ttttacgttt ctcgttcagc tttcttgtac aaagtggtta
     8461 ccggtggagt gcaggtggaa accatctccc caggagacgg gcgcaccttc cccaagcgcg
     8521 gccagacctg cgtggtgcac tacaccggga tgcttgaaga tggaaagaaa gttgattcct
     8581 cccgggacag aaacaagccc tttaagttta tgctaggcaa gcaggaggtg atccgaggct
     8641 gggaagaagg ggttgcccag atgagtgtgg gtcagagagc caaactgact atatctccag
     8701 attatgccta tggtgccact gggcacccag gcatcatccc accacatgcc actctcgtct
     8761 tcgatgtgga gcttctaaaa ctggaaggcg gctaccccta cgacgtgccc gactacgccg
     8821 gctatccgta tgatgtcccg gactatgcag gctaaacgcg ttaagtcgac aatcaacctc
     8881 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     8941 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     9001 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     9061 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     9121 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     9181 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     9241 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     9301 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     9361 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     9421 ctcagacgag tcggatctcc ctttgggccg cctccccgcg tcgactttaa gaccaatgac
     9481 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     9541 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     9601 ccagatctga gcctgggagc t
//
