LOCUS       sequence-261761-        7658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA tagged ADAM10 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7658)
  AUTHORS   Gschwind A, Hart S, Fischer OM, Ullrich A
  TITLE     TACE cleavage of proamphiregulin regulates GPCR-induced 
            proliferation and motility of cancer cells.
  JOURNAL   EMBO J. 2003 May 15;22(10):2411-21.
  PUBMED    12743035
REFERENCE   2  (bases 1 to 7658)
  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: "EMBO J."; 
            date: "2003-05-15"; volume: "22(10)"; pages: "2411-21"
COMMENT     Sequence Label: pcDNA3-ADAM10-HA
FEATURES             Location/Qualifiers
     source          1..7658
                     /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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2929..2955
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2978..2995)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3015..3032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3021..3245
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3291..3719
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3378..3397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3588..3609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3728..3748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3733..4062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3913..4048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3975..3994
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4129..4923
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4183..4202)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4793..4812
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5097..5218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5134..5153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5188..5207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5267..5283)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5267..5283)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5280..5302)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5291..5307
                     /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(5315..5321,5322..5339,5340..5345))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5360..5381
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5498..5515)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5669..6257)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5749..5768)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6428..7219,7220..7288))
                     /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     7051..7070
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7289..7393)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7468..7487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgcca ccatggtgtt gctgagagtg ttaattctgc tcctctcctg ggcggcgggg
      721 atgggaggtc agtatgggaa tcctttaaat aaatatatca gacattatga aggattatct
      781 tacaatgtgg attcattaca ccaaaaacac cagcgtgcca aaagagcagt ctcacatgaa
      841 gaccaatttt cacgtctaga tttccatgcc catggaagac atttcaacct acgaatgaag
      901 agggacactt cccttttcag tgatgaattt aaagtagaaa catcaaataa agtacttggt
      961 tatgatacct ctcatattta cactggacat atttatggtg aagaaggaag ttttggccat
     1021 gggtctgtta ttgatggaag atttgaagga ttcatccaga ctcgtggtgg cacattttat
     1081 gttgagccag cagagagata tattaaagac cgaactctgc catttcactc tgtcatttat
     1141 catgaagatg atattaacta tccccataaa tacggtcctc aggggggctg tgcagatcat
     1201 tcagtatttg aaagaatgag gaaataccag atgactggtg tagaggaagt aacacagata
     1261 cctcaagaag aacatgctgc taatggtcca gaacttctga ggaaaaaacg tacaacttca
     1321 gctgaaaaaa atacttgtca gctttatatt cagactgatc atttgttctt taaatattac
     1381 ggaacacgag aagctgtgat tgcccagata tccagtcatg ttaaagcgat tgatacaatt
     1441 taccagacca cagacttctc cggaatccgt aacatcagtt tcatggtgaa acgcataaga
     1501 atcaatacaa ctgctgatga gaaggaccct acgaatcctt tccgtttccc aaatattggt
     1561 gtggagaagt ttctggaatt gaattctgag cagaatcatg atgactactg tttggcccat
     1621 gtcttcacag accgagattt tgatgatggc gtacttggtc tggcttgggt tggagcacct
     1681 tcaggaagct ctggaggaat atgtgaaaaa agtaaactct attcagatgg taagaagaag
     1741 tccttaaaca ctggaattat tactgttcag aactatgggt ctcatgtacc tcccaaagtc
     1801 tctcacatta cttttgctca cgaagttgga cataactttg gatccccaca tgattctgga
     1861 acagagtgca caccaggaga atctaagaat ttgggtcaaa aagaaaatgg caattacatc
     1921 atgtatgcaa gagcaacatc tggggacaaa cttaacaaca ataaattctc actctgtagt
     1981 attagaaata taagccaagt tcttgagaag aagagaaaca actgttttgt tgaatctggc
     2041 caacctattt gtggaaatgg aatggtagaa caaggtgaag aatgtgattg tggctatagt
     2101 gaccagtgta aagatgaatg ctgcttcgat gcaaatcaac cagagggaag aaaatgcaaa
     2161 ctgaaacctg ggaaacagtg cagtccaagt caaggtcctt gttgtacagc acagtgtgca
     2221 ttcaagtcaa agtctgagaa gtgtcgggat gattcagact gtgcaaggga aggaatatgt
     2281 aatggcttca cagctctctg cccagcatct gaccctaaac caaacttcac agactgtaat
     2341 aggcatacac aagtgtgcat taatgggcaa tgtgcaggtt ctatctgtga gaaatatggc
     2401 ttagaggagt gtacgtgtgc cagttctgat ggcaaagatg ataaagaatt atgccatgta
     2461 tgctgtatga agaaaatgga cccatcaact tgtgccagta cagggtctgt gcagtggagt
     2521 aggcacttca gtggtcgaac catcaccctg caacctggat ccccttgcaa cgattttaga
     2581 ggttactgtg atgtcttcat gcggtgcaga ttagtagatg ctgatggtcc tctagctagg
     2641 cttaaaaaag caatttttag tccagagctc tatgaaaaca ttgctgaatg gattgtggct
     2701 cattggtggg cagtattact tatgggaatt gctctgatca tgctaatggc tggatttatt
     2761 aagatatgcg gtgttcatac tccaagtagt aatccaaagc tgcctcctcc taaaccactt
     2821 ccaggcactt taaagaggag gagacctcca cagcccattc agcaacccca gcgtcagcgg
     2881 ccccgagaga gttatcaaat gggacacatg agacgcggag gagggcccta cccatacgat
     2941 gttccagatt acgcttaact cgagctagcg gccgcggcta tagtgtcacc taaatgctag
     3001 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3061 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3121 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3181 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3241 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3301 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3361 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3421 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3481 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3541 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3601 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3661 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3721 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3781 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3841 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3901 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3961 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4021 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4081 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4141 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4201 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4261 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4321 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4381 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4441 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4501 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4561 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4621 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4681 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     4741 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     4801 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     4861 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4921 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4981 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5041 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5221 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5281 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5341 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5401 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5461 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5521 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5581 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5641 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5701 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5761 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5821 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5881 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5941 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6001 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6061 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6121 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6181 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6241 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6301 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6361 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6421 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6481 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6541 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6601 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6661 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6721 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6781 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6841 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6901 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6961 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7021 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7081 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7141 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7201 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7261 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7321 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7381 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7441 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7501 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7561 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     7621 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
