LOCUS       .                       9410 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             5406..5801
                     /label="BSD"
                     /note="blasticidin S deaminase"
     misc_feature    5847..5964
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6019..6522
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    6543..6667
                     /label="attR1"
                     /note="recombination site for the Gateway® LR reaction"
     promoter        6692..6722
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             6776..7432
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             7777..8079
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(8123..8247)
                     /label="attR2"
                     /note="recombination site for the Gateway® LR reaction"
     CDS             8255..8575
                     /label="DmrB"
                     /note="F36V mutant of FK506-binding protein FKBP12"
     CDS             8582..8608
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             8612..8638
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    8660..9248
                     /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 agaggtaccg
     5401 ccaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     5461 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     5521 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     5581 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     5641 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     5701 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     5761 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaacgaatt ctagatcttg
     5821 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     5881 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     5941 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccactttgg
     6001 cgccggctcg aggcctgcag gttgcgcctt ttccaaggca gccctgggtt tgcgcaggga
     6061 cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg ccgaccctgg gtctcgcaca
     6121 ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg ctacccttgt gggccccccg
     6181 gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct tgcggttcgc ggcgtgccgg
     6241 acgtgacaaa cggaagccgc acgtctcact agtaccctcg cagacggaca gcgccaggga
     6301 gcaatggcag cgcgccgacc gcgatgggct gtggccaata gcggctgctc agcagggcgc
     6361 gccgagagca gcggccggga aggggcggtg cgggaggcgg ggtgtggggc ggtagtgtgg
     6421 gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc ctccggagcg cacgtcggca
     6481 gtcggctccc tcgttgaccg aatcaccgac ctctctcccc aggggctagc atcgattgat
     6541 caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
     6601 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     6661 cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa
     6721 tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga
     6781 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     6841 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     6901 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     6961 tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct
     7021 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     7081 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     7141 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     7201 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     7261 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     7321 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     7381 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaagatc
     7441 tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg
     7501 tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag
     7561 cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg
     7621 tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg
     7681 aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga
     7741 acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat
     7801 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc
     7861 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     7921 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     7981 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     8041 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata
     8101 cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt
     8161 agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc
     8221 tcgttcagct ttcttgtaca aagtggttac cggtggagtg caggtggaaa ccatctcccc
     8281 aggagacggg cgcaccttcc ccaagcgcgg ccagacctgc gtggtgcact acaccgggat
     8341 gcttgaagat ggaaagaaag ttgattcctc ccgggacaga aacaagccct ttaagtttat
     8401 gctaggcaag caggaggtga tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg
     8461 tcagagagcc aaactgacta tatctccaga ttatgcctat ggtgccactg ggcacccagg
     8521 catcatccca ccacatgcca ctctcgtctt cgatgtggag cttctaaaac tggaaggcgg
     8581 ctacccctac gacgtgcccg actacgccgg ctatccgtat gatgtcccgg actatgcagg
     8641 ctaaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     8701 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     8761 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     8821 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     8881 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     8941 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     9001 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     9061 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     9121 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     9181 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     9241 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     9301 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     9361 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct
//
