LOCUS       sequence-128132-        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-WW mutant
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             625..648
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2172..2794
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2371..2390)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2818..2838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2823..3152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3003..3138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3190..3209)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3191..3209)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3223..3239)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3223..3239)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3236..3258)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3247..3263
                     /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(3271..3277,3278..3295,3296..3301))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3316..3337
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3454..3471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3625..4213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3705..3724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4384..5175,5176..5244))
                     /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     5007..5026
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5245..5349)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5376..5831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5463..5482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5673..5694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5957..5979
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5971..5988
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5972..5988
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      601 aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct tgcggccgcg
      661 aattcaagct tagccaccat ggactacaaa gacgatgacg ataaagcaag gctcgaatcg
      721 gtacctaagg atcccgggca gcagccgccg cctcaaccgg ccccccaggg ccaagggcag
      781 ccgccttcgc agcccccgca ggggcagggc ccgccgtccg gacccgggca accggcaccc
      841 gcggcgaccc aggcggcgcc gcaggcaccc cccgccgggc atcagatcgt gcacgtccgc
      901 ggggactcgg agaccgacct ggaggcgctc ttcaacgccg tcatgaaccc caagacggcc
      961 aacgtgcccc agaccgtgcc catgaggctc cggaagctgc ccgactcctt cttcaagccg
     1021 ccggagccca aatcccactc ccgacaggcc agtactgatg caggcactgc aggagccctg
     1081 actccacagc atgttcgagc tcattcctct ccagcttctc tgcagttggg agctgtttct
     1141 cctgggacac tgacccccac tggagtagtc tctggcccag cagctacacc cacagctcag
     1201 catcttcgac agtcttcttt tgagatacct gatgatgtac ctctgccagc aggttgggag
     1261 atggcaaaga catcttctgg tcagagatac ttcttaaatc acatcgatca gacaacaaca
     1321 gcgcaggacg ccaggaaggc catgctgtcc cagatgaacg tcacagcccc caccagtcca
     1381 ccagtgcagc agaatatgat gaactcggct tcagccatga accagagaat cagtcagagt
     1441 gctccagtga aacagccacc acccctggct ccccagagcc cacagggagg cgtcatgggt
     1501 ggcagcaact ccaaccagca gcaacagatg cgactgcagc aactgcagat ggagaaggag
     1561 aggctgcggc tgaaacagca agaactgctt cggcaggtga ggccacagga gttagccctg
     1621 cgtagccagt taccaacact ggagcaggat ggtgggactc aaaatccagt gtcttctccc
     1681 gggatgtctc aggaattgag aacaatgacg accaatagct cagatccttt ccttaacagt
     1741 ggcacctatc actctcgaga tgagagtaca gacagtggac taagcatgag cagctacagt
     1801 gtccctcgaa ccccagatga cttcctgaac agtgtggatg agatggatac aggtgatact
     1861 atcaaccaaa gcaccctgcc ctcacagcag aaccgtttcc cagactacct tgaagccatt
     1921 cctgggacaa atgtggacct tggaacactg gaaggagatg gaatgaacat agaaggagag
     1981 gagctgatgc caagtctgca ggaagctttg agttctgaca tccttaatga catggagtct
     2041 gttttggctg ccaccaagct agataaagaa agctttctta catggttata gagccctcag
     2101 gcagactgaa ttatctgcag aattctgcag atatccagca cagtggcggc cgctcgagtc
     2161 tagaggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     2221 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2281 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     2341 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     2401 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     2461 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     2521 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     2581 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     2641 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     2701 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     2761 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga
     2821 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2881 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2941 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3001 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3061 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3121 gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga
     3181 aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt
     3241 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3301 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3361 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3421 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3481 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3541 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3601 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3661 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3721 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3781 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3841 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3901 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3961 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4021 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4081 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4141 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4201 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4261 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4321 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4381 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4441 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4501 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4561 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4621 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4681 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4741 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4801 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4861 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4921 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4981 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5041 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5101 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5161 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5221 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5281 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5341 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     5401 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5461 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5521 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5581 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5641 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5701 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5761 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5821 gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5881 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5941 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc
     6001 tatacattga atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc
     6061 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     6121 ggctcatgtc caatatgacc g
//
