LOCUS       V040306                 8673 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040306
VERSION     V040306
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     protein_bind    113..134
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        149..179
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    187..203
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /label="lac operator"
     primer_bind     211..227
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     promoter        248..266
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /label="T3 promoter"
     promoter        294..520
                     /note="Rous sarcoma virus enhancer/promoter"
                     /label="RSV promoter"
     LTR             521..701
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    748..873
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1366..1599
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1784..1828
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    2095..2211
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     enhancer        2234..2537
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2538..2741
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2832..3680
                     /label="KLF6(NM_001300)"
                     /note="KLF6(NM_001300)"
                     /gene="KLF6"
     CDS             3681..3770
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     promoter        3805..4134
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4144..4743
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    4810..5398
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5485..5718
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5790..5911
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5951..6086
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6107..6125)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6135..6151)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6293..6748
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6774..6878
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             6879..7739
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      7910..8498
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
ORIGIN
        1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
       61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      241 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc
      301 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca
      361 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt
      421 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg
      481 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca
      541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
      601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
      661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac
      721 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg
      781 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga
      841 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg
      901 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata
      961 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca
     1021 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa
     1081 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata
     1141 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc
     1201 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag
     1261 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa
     1321 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct
     1381 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca
     1441 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga
     1501 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat
     1561 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg
     1621 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga
     1681 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag
     1741 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt
     1801 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg
     1861 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc
     1921 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca
     1981 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag
     2041 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa
     2101 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
     2161 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag
     2221 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2521 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     2581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     2641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     2701 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     2761 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagacgg
     2821 gatccgccac catggacgtg ctccccatgt gcagcatctt ccaggagctc cagatcgtgc
     2881 acgagaccgg ctacttctcg gcgctgccgt ctctggagga gtactggcaa cagacctgcc
     2941 tagagctgga acgttacctc cagagcgagc cctgctatgt ttcagcctca gaaatcaaat
     3001 ttgacagcca ggaagatctg tggaccaaaa tcattctggc tcgggagaaa aaggaggaat
     3061 ccgaactgaa gatatcttcc agtcctccag aggacactct catcagcccg agcttttgtt
     3121 acaacttaga gaccaacagc ctgaactcag atgtcagcag cgaatcctct gacagctccg
     3181 aggaactttc tcccacggcc aagtttacct ccgaccccat tggcgaagtt ttggtcagct
     3241 cgggaaaatt gagctcctct gtcacctcca cgcctccatc ttctccggaa ctgagcaggg
     3301 aaccttctca actgtggggt tgcgtgcccg gggagctgcc ctcgccaggg aaggtgcgca
     3361 gcgggacttc ggggaagcca ggtgacaagg gaaatggcga tgcctccccc gacggcagga
     3421 ggagggtgca ccggtgccac tttaacggct gcaggaaagt ttacaccaaa agctcccact
     3481 tgaaagcaca ccagcggacg cacacaggag aaaagcctta cagatgctca tgggaagggt
     3541 gtgagtggcg ttttgcaaga agtgatgagt taaccaggca cttccgaaag cacaccgggg
     3601 ccaagccttt taaatgctcc cactgtgaca ggtgtttttc caggtctgac cacctggccc
     3661 tgcacatgaa gaggcacctc gaacaaaaac taatatcaga ggaagatttg gaacagaaac
     3721 taatcagtga agaagattta gagcaaaagt taatttccga ggaggactta tagacgcgtc
     3781 gaccagtggt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3901 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3961 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4021 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4081 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     4141 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4201 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4261 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4321 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4381 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4441 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4501 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4561 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4621 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4681 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     4741 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     4801 ccagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4861 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4921 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     4981 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5041 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5101 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5161 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     5221 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5281 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5341 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     5401 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5461 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     5521 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     5581 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     5641 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     5701 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     5761 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     5821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5881 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     5941 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6001 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6061 gtagtgagga ggcttttttg gaggcctagg gacgtaccca attcgcccta tagtgagtcg
     6121 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     6181 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     6241 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     6301 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     6361 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     6421 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     6481 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     6541 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6601 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     6661 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6721 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     6781 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6841 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6901 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6961 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     7021 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     7081 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     7141 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     7201 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     7261 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     7321 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7381 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7441 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7501 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7561 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7621 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7681 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7741 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7801 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7861 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7921 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7981 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     8041 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     8101 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     8161 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     8221 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     8281 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     8341 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8401 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8461 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8521 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     8581 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     8641 cgaccgagcg cagcgagtca gtgagcgagg aag
//
