LOCUS       sequence-340319-        7128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encoding dog Hbegf..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  AUTHORS   Lin S, Hirayama D, Maryu G, Matsuda K, Hino N, Deguchi E, Aoki K, 
            Iwamoto R, Terai K, Matsuda M
  TITLE     Redundant roles of EGFR ligands in the ERK activation waves during 
            collective cell migration.
  JOURNAL   Life Sci Alliance. 2021 Oct 19;5(1). pii: 5/1/e202101206. doi: 
            10.26508/lsa.202101206. Print 2022 Jan.
  PUBMED    34667080
REFERENCE   2  (bases 1 to 7128)
  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: "Life Sci 
            Alliance."; date: "2021-10-19"; volume: "5(1). pii: 5/1/e202101206. 
            doi"; pages: " 10.26508/lsa.202101206. Print 2022 Jan"
COMMENT     Sequence Label: pPBpuro-UbC-canisHbegf
FEATURES             Location/Qualifiers
     source          1..7128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     259..278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     misc_feature    1241..1790
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1385..1402)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1612..1631
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1793..2392
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1793..1812)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2289..2309
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(2419..2436)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2425..2649
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2696..2712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2720..2736
                     /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(2744..2750,2751..2768,2769..2774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2789..2810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     repeat_region   3044..3106
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(3147..3167)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3147..3165)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3186..3202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3210..3226
                     /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(3234..3240,3241..3258,3259..3264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3279..3300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3417..3434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3588..4176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3668..3687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4347..5138,5139..5207))
                     /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     4970..4989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5208..5312)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5338..5793)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5475..5496)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5687..5706
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5920..5942
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5934..5951
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5935..5951
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5961..5980
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5961..5979
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   6009..6043
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        join(6339..7128,1..422)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
       61 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
      121 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
      181 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctctggtcg gttttatgta
      241 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
      301 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
      361 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
      421 acggatccat gaattcacca tgaagctgct gcggtcagtg gtgctgaagc tctttctggc
      481 tgcagtgctc tcggcgtcgg tgactggcga gagcttgggg cgtcttcgga gagggctggc
      541 ggccggaact ggcaacccgg actctcccac cgaatccacg gaccggctgc tgcccccgga
      601 aggcggccgg gccagggaag tcctggactt agaagagacg gacctggacc ttttaagagc
      661 tgctttctcc tccaagccac aggctctggc cacacccagt aaggaggaac gtgggaaaaa
      721 aaagaagaaa ggcaaaggct tagggaggaa gagagacccg tgtcttcgga aatacaagga
      781 cttctgcatc catggagaat gcaaatatgt gaaggagctc cgggctccat cctgcatctg
      841 ccaccccggt taccatggag agaggtgcca tgggctgagc cttccagtag aaaatcgctt
      901 atatacttac gaccatacaa ccatcttggc tgtggtggct gtggtgctgt catctgtctg
      961 tctgcttgtc atcgtggggc ttctcatgtt taggtaccat aggagaggag gttatgatgt
     1021 ggaaagtgaa gagaaagtga agttaggcat gactacttcc cactgagaga gacctgtact
     1081 caaggaatct gcagggggac tgctacctcc gagaagatac agctgatcat agactctgca
     1141 gagggaaagg acgtcacagc taggcggccg ctctagagtc aactaactta agctagcaac
     1201 ggtttccctc tagcgggatc aattccgccc ccccccccta acgttactgg ccgaagccgc
     1261 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     1321 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     1381 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     1441 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     1501 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     1561 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     1621 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     1681 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     1741 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgataata ccatgaccga
     1801 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     1861 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     1921 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     1981 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     2041 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     2101 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     2161 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     2221 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     2281 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     2341 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gatcgatgat
     2401 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2461 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2521 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2581 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2641 gctctatggc ttctgaggcg gaaagaacct gcagcccaag cttggcgtaa tcatggtcat
     2701 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2761 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2821 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatctta aaagttttgt
     2881 tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata
     2941 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
     3001 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
     3061 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagtatac
     3121 gcgtttaatt aatccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     3181 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3241 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3301 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3361 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3421 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3481 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3781 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4021 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4201 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4381 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4441 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4501 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4561 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4621 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4681 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4741 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4801 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4861 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4921 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4981 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5041 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5101 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5161 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5221 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5281 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5341 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5401 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5461 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     5521 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     5581 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     5641 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     5701 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     5761 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     5821 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     5881 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
     5941 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggg cgcgccattc
     6001 tagattaacc ctagaaagat agtctgcgta aaattgacgc atgcattctt gaaatattgc
     6061 tctctctttc taaatagcgc gaatccgtcg ctgtgcattt aggacatctc agtcgccgct
     6121 tggagctccc gtgaggcgtg cttgtcaatg cggtaagtgt cactgatttt gaactataac
     6181 gaccgcgtga gtcaaaatga cgcatgatta tcttttacgt gacttttaag atttaactca
     6241 tacgataatt atattgttat ttcatgttct acttacgtga taacttatta tatatatatt
     6301 ttcttgttat agatatcaac tagaatgcta gcctcgaggg cctccgcgcc gggttttggc
     6361 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
     6421 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     6481 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     6541 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     6601 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     6661 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cagttcttgt ttgtggatcg
     6721 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     6781 gggccgctcg gtgggacgga ggcgtgtgga gagaccgcca agggctgtag tctgggtccg
     6841 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     6901 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     6961 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     7021 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     7081 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtg
//
