LOCUS       .                       9650 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        13..516
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             525..848
                     /label="FKBP-V"
                     /note="F36V mutant of FK506-binding protein FKBP12"
     CDS             855..881
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             885..911
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     protein_bind    927..1051
                     /label="attR1"
                     /note="recombination site for the Gateway® LR reaction"
     promoter        1076..1106
                     /label="lac UV5 promoter"
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             1160..1816
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     CDS             2161..2463
                     /label="ccdB"
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(2507..2631)
                     /label="attR2"
                     /note="recombination site for the Gateway® LR reaction"
     misc_feature    2683..3271
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3343..3576
                     /label="3' LTR (ΔU3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3654..3788
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(3971..3989)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3999..4015)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      4157..4612
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4638..4742
                     /label="AmpR promoter"
     CDS             4743..5600
                     /label="AmpR"
                     /note="β-lactamase"
     rep_origin      5774..6362
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    6650..6671
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6686..6716
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6724..6740
                     /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     6748..6764
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        6785..6803
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6831..7057
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     misc_feature    7285..7410
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7903..8136
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8321..8365
                     /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        8596..8806
                     /label="SV40 promoter"
                     /note="SV40 early promoter"
     CDS             8846..9442
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    9491..9608
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
ORIGIN
        1 ctcgaggcct gcaggttgcg ccttttccaa ggcagccctg ggtttgcgca gggacgcggc
       61 tgctctgggc gtggttccgg gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc
      121 acgtccgttc gcagcgtcac ccggatcttc gccgctaccc ttgtgggccc cccggcgacg
      181 cttcctgctc cgcccctaag tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga
      241 caaacggaag ccgcacgtct cactagtacc ctcgcagacg gacagcgcca gggagcaatg
      301 gcagcgcgcc gaccgcgatg ggctgtggcc aatagcggct gctcagcagg gcgcgccgag
      361 agcagcggcc gggaaggggc ggtgcgggag gcggggtgtg gggcggtagt gtgggccctg
      421 ttcctgcccg cgcggtgttc cgcattctgc aagcctccgg agcgcacgtc ggcagtcggc
      481 tccctcgttg accgaatcac cgacctctct ccccaggggc tagcatggga gtgcaggtgg
      541 aaaccatctc cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc
      601 actacaccgg gatgcttgaa gatggaaaga aagttgattc ctcccgggac agaaacaagc
      661 cctttaagtt tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc
      721 agatgagtgt gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca
      781 ctgggcaccc aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa
      841 aactggaagg cggctacccc tacgacgtgc ccgactacgc cggctatccg tatgatgtcc
      901 cggactatgc aggcatcgat agatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt
      961 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     1021 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
     1081 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc
     1141 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     1201 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     1261 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     1321 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     1381 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     1441 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     1501 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     1561 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     1621 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     1681 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     1741 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     1801 gtggcagggc ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc
     1861 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     1921 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     1981 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     2041 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     2101 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
     2161 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     2221 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     2281 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     2341 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     2401 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     2461 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
     2521 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     2581 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg ttaccggtta
     2641 gtaatgaact agttaagtcg acaatcaacg cgttaagtcg acaatcaacc tctggattac
     2701 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2761 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2821 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2881 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2941 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3001 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3061 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     3121 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3181 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3241 agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg
     3301 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     3361 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     3421 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     3481 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     3541 tcagaccctt ttagtcagtg tggaaaatct ctagcagtac gtatagtagt tcatgtcatc
     3601 ttattattca gtatttataa cttgcaaaga aatgaatatc agagagtgag aggaacttgt
     3661 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3721 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3781 tctggctcta gctatcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3841 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3901 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc tagggacgta
     3961 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     4021 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4081 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4141 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4201 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4261 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4321 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4381 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4441 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4501 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4561 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4621 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4681 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4741 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4801 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4861 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4921 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4981 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5041 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5101 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5161 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5221 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5281 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5341 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5401 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5461 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5521 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5581 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5641 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5701 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5881 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6421 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6481 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6541 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6601 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6661 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6721 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6781 gcgcaattaa ccctcactaa agggaacaaa agctggagct gcaagcttaa tgtagtctta
     6841 tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg ccttacaagg
     6901 agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc gtgccttatt
     6961 aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc cgcattgcag
     7021 agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt tagaccagat
     7081 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     7141 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     7201 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     7261 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     7321 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     7381 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7441 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7501 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7561 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7621 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7681 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7741 cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     7801 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     7861 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     7921 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     7981 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     8041 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     8101 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     8161 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     8221 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     8281 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     8341 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     8401 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     8461 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     8521 cccaaccccg aggggaccca tgcattgcat ctcaattagt cagcaaccag gtgtggaaag
     8581 tccccaggct ccccagcagg cagaagtatg caaagcatgc gtctcaatta gtcagcaacc
     8641 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     8701 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     8761 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     8821 tctagaggta ccaccggagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac
     8881 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     8941 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     9001 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     9061 cgtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     9121 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     9181 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     9241 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     9301 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     9361 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     9421 catgacccgc aagcccggtg ccagttaacg aattctagat cttgagacaa atggcagtat
     9481 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     9541 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     9601 aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg
//
