LOCUS       V027897                 8828 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027897
VERSION     V027897
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             2824..3879
                     /label="RAD51AP1(NM_001130862)"
                     /note="RAD51AP1(NM_001130862)"
                     /gene="RAD51AP1"
     CDS             3886..3909
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        3960..4289
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4299..4898
                     /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    4965..5553
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5640..5873
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5945..6066
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6106..6241
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        complement(6262..6280)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(6290..6306)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     rep_origin      6448..6903
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6929..7033
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7034..7894
                     /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      8065..8653
                     /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 ctctagagcc
     2821 accatggtgc ggcctgtgag acataagaaa ccagtcaatt actcacagtt tgaccactct
     2881 gacagtgatg atgattttgt ttctgcaact gtacctttaa acaagaaatc cagaacagca
     2941 ccaaaggagt taaaacaaga taaaccaaaa cctaacttga acaatctccg gaaagaagaa
     3001 atcccagtac aagagaaaac ccctaaaaaa agactccctg aaggtacttt tagtattcca
     3061 gctagtgcag tgccttgtac aaagatggct ttagatgaca agctctacca gagagactta
     3121 gaagttgcac tagctttatc agtgaaggaa cttccaacag tcaccactaa tgtgcagaac
     3181 tctcaagata aaagcattga aaaacatggc agtagtaaaa tagaaacaat gaataagtct
     3241 cctcatatct ctaattgcag tgtagccagt gattatttag atttggataa gattactgtg
     3301 gaagatgatg ttggtggtgt tcaagggaaa agaaaagcag catctaaagc tgcagcacag
     3361 cagaggaaga ttcttctgga aggcagtgat ggtgatagtg ctaatgacac tgaaccagac
     3421 tttgcacctg gtgaagattc tgaggatgat tctgattttt gtgagagtga ggataatgac
     3481 gaagacttct ctatgagaaa aagtaaagtt aaagaaatta aaaagaaaga agtgaaggta
     3541 aaatccccag tagaaaagaa agagaagaaa tctaaatcca aatgtaatgc tttggtgact
     3601 tcggtggact ctgctccagc tgccgtcaaa tcagaatctc agtccttgcc aaaaaaggtt
     3661 tctctgtctt cagataccac taggaaacca ttagaaatac gcagtccttc agctgaaagc
     3721 aagaaaccta aatgggtccc accagcggca tctggaggta gcagaagtag cagcagccca
     3781 ctggtggtag tgtctgtgaa gtctcccaat cagagtctcc gccttggctt gtccagatta
     3841 gcacgagtta aacctttgca tccaaatgcc actagcaccg aattcgatta caaggatgac
     3901 gacgataagt gaggatccgg actagttaac gcgtcgacca gtggtcgacc ctgtggaatg
     3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac aagcccacgg
     4321 tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg
     4381 ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg
     4441 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     4501 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     4561 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     4621 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4681 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4741 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4801 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4861 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc
     4921 gcccgaccga aaggagcgca cgaccccatg catcgccagt cgacaatcaa cctctggatt
     4981 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     5041 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     5101 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     5161 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     5221 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     5281 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     5341 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     5401 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     5461 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     5521 cgagtcggat ctccctttgg gccgcctccc cgcctggaat tcgagctcgg tacctttaag
     5581 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     5641 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     5701 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     5761 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5821 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     5881 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     5941 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6001 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6061 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     6121 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6181 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6241 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     6301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     6361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     6421 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     6481 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     6541 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     6601 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     6661 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     6721 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     6781 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6841 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     6901 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     7021 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     7081 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     7141 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     7201 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     7261 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7321 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7381 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7441 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7501 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7561 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7621 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7681 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7741 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7801 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7861 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7921 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7981 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     8041 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     8101 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     8161 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     8221 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     8281 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     8341 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     8401 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     8461 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     8521 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8581 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8641 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8701 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     8761 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     8821 cgaggaag
//
