LOCUS       sequence-323905-        6141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   Komuro A, Nagai M, Navin NE, Sudol M
  TITLE     WW domain-containing protein YAP associates with ErbB-4 and acts as 
            a co-transcriptional activator for the carboxyl-terminal fragment of
            ErbB-4 that translocates to the nucleus.
  JOURNAL   J Biol Chem. 2003 Aug 29. 278(35):33334-41.
  PUBMED    12807903
REFERENCE   2  (bases 1 to 6141)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2003 Aug 29. 278(35):33334-41."
COMMENT     Sequence Label: p2xFLAGhYAP1
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..641
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             675..698
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2165..2787
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2364..2383)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2811..2831)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2816..3145
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2996..3131
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3183..3202)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3184..3202)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3216..3232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3216..3232)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3229..3251)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3240..3256
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3264..3270,3271..3288,3289..3294))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3309..3330
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3447..3464)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3618..4206)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3698..3717)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4377..5168,5169..5237))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5000..5019
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5238..5342)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5369..5824
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5666..5687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5950..5972
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5964..5981
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5965..5981
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggac tacaaagacg atgacgacaa gcttgcggcc gcgaattcaa
      661 gcttagccac catggactac aaagacgatg acgataaagc aaggctcgaa tcggtaccta
      721 aggatcccgg gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt
      781 cgcagccccc gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga
      841 cccaggcggc gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact
      901 cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc
      961 cccagaccgt gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc
     1021 ccaaatccca ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac
     1081 agcatgttcg agctcattcc tctccagctt ctctgcagtt gggagctgtt tctcctggga
     1141 cactgacccc cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc
     1201 gacagtcttc ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa
     1261 agacatcttc tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg
     1321 accccaggaa ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc
     1381 agcagaatat gatgaactcg gcttcagcca tgaaccagag aatcagtcag agtgctccag
     1441 tgaaacagcc accacccctg gctccccaga gcccacaggg aggcgtcatg ggtggcagca
     1501 actccaacca gcagcaacag atgcgactgc agcaactgca gatggagaag gagaggctgc
     1561 ggctgaaaca gcaagaactg cttcggcagg tgaggccaca ggagttagcc ctgcgtagcc
     1621 agttaccaac actggagcag gatggtggga ctcaaaatcc agtgtcttct cccgggatgt
     1681 ctcaggaatt gagaacaatg acgaccaata gctcagatcc tttccttaac agtggcacct
     1741 atcactctcg agatgagagt acagacagtg gactaagcat gagcagctac agtgtccctc
     1801 gaaccccaga tgacttcctg aacagtgtgg atgagatgga tacaggtgat actatcaacc
     1861 aaagcaccct gccctcacag cagaaccgtt tcccagacta ccttgaagcc attcctggga
     1921 caaatgtgga ccttggaaca ctggaaggag atggaatgaa catagaagga gaggagctga
     1981 tgccaagtct gcaggaagct ttgagttctg acatccttaa tgacatggag tctgttttgg
     2041 ctgccaccaa gctagataaa gaaagctttc ttacatggtt atagagccct caggcagact
     2101 gaattatctg cagaattctg cagatatcca gcacagtggc ggccgctcga gtctagagga
     2161 tcccgggtgg catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac
     2221 tccagtgccc accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt
     2281 gtccttctat aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa
     2341 gacaacctgt agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc
     2401 ttggctcact gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga
     2461 gttgttggga ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag
     2521 acggggtttc accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc
     2581 accttggcct cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct
     2641 tctgatttta aaataactat accagcagga ggacgtccag acacagcata ggctacctgg
     2701 ccatgcccaa ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg
     2761 gtccactcag tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg
     2821 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2881 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2941 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3001 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3061 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3121 ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa
     3181 ttccctatag tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat
     3241 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3301 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3361 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3421 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3481 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3541 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3601 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3661 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3721 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3781 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3841 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3901 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3961 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4021 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4081 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4141 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4201 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4261 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4321 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4381 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4441 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4501 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4561 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4621 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4681 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4741 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4801 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4861 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4921 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4981 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5041 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5101 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5161 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5221 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5281 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5341 cgcacatttc cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5821 atttccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     5881 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     5941 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctg atctatacat
     6001 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     6061 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     6121 gtccaatatg accgccatgt t
//
