LOCUS       sequence-176437-        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-S127A
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..63
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     55..72
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     56..72
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        233..612
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        613..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     767..787
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             850..873
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             907..930
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2397..3019
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2596..2615)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3043..3063)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3048..3377
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3228..3363
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3415..3434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3416..3434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3448..3464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3448..3464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3461..3483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3472..3488
                     /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(3496..3502,3503..3520,3521..3526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3541..3562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3679..3696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3850..4438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3930..3949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4609..5400,5401..5469))
                     /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     5232..5251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5470..5574)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5601..6056
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5688..5707)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5898..5919
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
       61 acgacggcca gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta
      121 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
      181 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga
      241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      481 catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      541 gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc
      601 gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac
      661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      721 aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt
      781 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaattga
      841 tctaccatgg actacaaaga cgatgacgac aagcttgcgg ccgcgaattc aagcttagcc
      901 accatggact acaaagacga tgacgataaa gcaaggctcg aatcggtacc taaggatccc
      961 gggcagcagc cgccgcctca accggccccc cagggccaag ggcagccgcc ttcgcagccc
     1021 ccgcaggggc agggcccgcc gtccggaccc gggcaaccgg cacccgcggc gacccaggcg
     1081 gcgccgcagg caccccccgc cgggcatcag atcgtgcacg tccgcgggga ctcggagacc
     1141 gacctggagg cgctcttcaa cgccgtcatg aaccccaaga cggccaacgt gccccagacc
     1201 gtgcccatga ggctccggaa gctgcccgac tccttcttca agccgccgga gcccaaatcc
     1261 cactcccgac aggccagtac tgatgcaggc actgcaggag ccctgactcc acagcatgtt
     1321 cgagctcatg cctctccagc ttctctgcag ttgggagctg tttctcctgg gacactgacc
     1381 cccactggag tagtctctgg cccagcagct acacccacag ctcagcatct tcgacagtct
     1441 tcttttgaga tacctgatga tgtacctctg ccagcaggtt gggagatggc aaagacatct
     1501 tctggtcaga gatacttctt aaatcacatc gatcagacaa caacatggca ggaccccagg
     1561 aaggccatgc tgtcccagat gaacgtcaca gcccccacca gtccaccagt gcagcagaat
     1621 atgatgaact cggcttcagc catgaaccag agaatcagtc agagtgctcc agtgaaacag
     1681 ccaccacccc tggctcccca gagcccacag ggaggcgtca tgggtggcag caactccaac
     1741 cagcagcaac agatgcgact gcagcaactg cagatggaga aggagaggct gcggctgaaa
     1801 cagcaagaac tgcttcggca ggtgaggcca caggagttag ccctgcgtag ccagttacca
     1861 acactggagc aggatggtgg gactcaaaat ccagtgtctt ctcccgggat gtctcaggaa
     1921 ttgagaacaa tgacgaccaa tagctcagat cctttcctta acagtggcac ctatcactct
     1981 cgagatgaga gtacagacag tggactaagc atgagcagct acagtgtccc tcgaacccca
     2041 gatgacttcc tgaacagtgt ggatgagatg gatacaggtg atactatcaa ccaaagcacc
     2101 ctgccctcac agcagaaccg tttcccagac taccttgaag ccattcctgg gacaaatgtg
     2161 gaccttggaa cactggaagg agatggaatg aacatagaag gagaggagct gatgccaagt
     2221 ctgcaggaag ctttgagttc tgacatcctt aatgacatgg agtctgtttt ggctgccacc
     2281 aagctagata aagaaagctt tcttacatgg ttatagagcc ctcaggcaga ctgaattatc
     2341 tgcagaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag gatcccgggt
     2401 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     2461 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     2521 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     2581 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     2641 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     2701 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     2761 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     2821 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     2881 taaaataact ataccagcag gaggacgtcc agacacagca taggctacct ggccatgccc
     2941 aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt gggtccactc
     3001 agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg tgtcagttag
     3061 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3121 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3181 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3241 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3301 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3361 gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt aattccctat
     3421 agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     3481 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3541 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3601 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3661 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3781 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3841 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3901 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3961 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4021 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4081 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4141 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4201 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4261 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4321 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4381 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4441 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4501 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4561 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4621 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4681 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4741 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4801 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4861 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4921 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4981 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5041 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5101 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5161 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5221 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5281 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5341 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5401 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5461 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5521 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5581 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5641 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5701 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5761 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5821 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5881 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5941 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     6001 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttccat
     6061 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6121 cgccagctgg cgaaaggggg a
//
