LOCUS       sequence-415332-        7935 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of HA tagged ADAM17 (TACE) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  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 7935)
  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-ADAM17-HA
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3440..3466
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3489..3506)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3526..3543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3532..3756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3802..4230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3889..3908)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4099..4120
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4239..4259)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4244..4573
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4424..4559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4486..4505
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4640..5434
                     /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(4694..4713)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5304..5323
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5608..5729
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5645..5664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5699..5718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5778..5794)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5778..5794)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5791..5813)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5802..5818
                     /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(5826..5832,5833..5850,5851..5856))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5871..5892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6009..6026)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6180..6768)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6260..6279)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6939..7730,7731..7799))
                     /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     7562..7581
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7800..7904)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcgccacca tgaggcagtc
      961 tctcctattc ctgaccagcg tggttccttt cgtgctgccg ccgcgacctc cggatgaccc
     1021 gggcttcggc ccccaccaga gactcgagaa gcttgattct ttgctctcag actacgatat
     1081 tctctcttta tctaatatcc agcagcattc ggtaagaaaa agagatctac agacttcaac
     1141 acatgtagaa acactactaa ctttttcagc tttgaaaagg cattttaaat tatacctgac
     1201 atcaagtact gaacgttttt cacaaaattt caaggtcgtg gtggtggatg gtaaaaacga
     1261 aagcgagtgc actgtaaaat ggcaggactt cttcactgga cacgtggttg gtgagcctga
     1321 ctctagggtt ctagcccaca taagagatga tgatgttata atcagaatca acacagatgg
     1381 ggccgaatat aacatagagc cactttggag atttgttaat gataccaaag acaaaagaat
     1441 gttagcttat aaatctgaag atatcaagaa tgtttcacgt ttgcagtctc caaaagtgtg
     1501 tggttattta aaagtggata atgaagagtt gctcccaaga gggttagtag acagagaacc
     1561 acctgaagag cttgttcatc gagtgaaaag aagagctgac ccagatccca tgaagaacac
     1621 gtgtaaatta ttggtggtag cagatcatcg cttctacaga tacatgggca gaggggagga
     1681 gagtacaact acaaattact taatagagct aattgacata gttgatgaca tctatcggaa
     1741 cacttcatgg gataatgcag gttttaaagg ctatggaata cagatagagc agattcgcat
     1801 tctcaagtct ccacaagagg taaaacctgg tgaaaagcac tacaacatgg caaaaagtta
     1861 cccaaatgaa gaaaaggatg cttgggatgt gaagatgttg ctagagcaat ttagctttga
     1921 tatagctgag gaagcatcta aagttagctt ggcacacctt ttcacatacc aagattttga
     1981 tatgggaact cttggattag cttatgttgg ctctcccaga gcaaacagcc atggaggtgt
     2041 ttgtccaaag gcttattata gcccagttgg gaagaaaaat atctatttga atagtggttt
     2101 gacgagcaca aagaattatg gtaaagccat ccttacaaag gaagctgacc tggttacaac
     2161 tcatgaattg ggacataatt ttggagcaga acatgatccg gatggtctag cagaatgtgc
     2221 cccgaatgag gaccagggag ggaaatatgt catgtatccc atagctgtga gtggcgatca
     2281 cgagaacaat aagatgttta caaactgcag taaacaatca atctataaga ccattgaaag
     2341 taaggcccag gagtgttttc aagaacgcag caataaagtt tgtgggaact cgagggtgga
     2401 tgaaggagaa gagtgtgatc ctggcatcat gtatctgaac aacgacacct gctgcaacag
     2461 cgactgcacg ttgaaggaag gtgtccagtg cagtgacagg aacagtcctt gctgtaaaaa
     2521 ctgtcagttt gagactgccc agaagaagtg ccaggaggcg attaatgcta cttgcaaagg
     2581 cgtgtcctac tgcacaggta atagcagtga gtgcccgcct ccaggaaatg ctgaagatga
     2641 cactgtttgc ttggatcttg gcaagtgtaa ggatgggaaa tgcatccctt tctgcgagtg
     2701 ggaacagcag ctggagtcct gtgcatgtag tgaaactgac aactcctgca aggtgtgctg
     2761 cagggacctt tctggccgct gtgtgcccta tgtcgatgct gaacaaaaga acttattttt
     2821 gaggaaagga aagccctgca cagtaggatt ttgtgacatg aatggcaaat gtgagaaacg
     2881 agtacaggat gtaattgaac gattttggga tttcattgac cagctgagca tcaatacttt
     2941 tggaaagttt ttagcagaca acatcgttgg gtctgtcctg gttttctcct tgatattttg
     3001 gattcctttc agcattcttg tccattgtgt ggataagaaa ttggataaac agtatgaatc
     3061 tctgtctctg tttcacccca gtaacgtcga aatgctgagc agcatggatt ctgcatcggt
     3121 tcgcattatc aaaccctttc ctgcgcccca gactccaggc cgcctgcagc ctgcccctgt
     3181 gatcccttcg gcgccagcag ctccaaaact ggaccaccag agaatggaca ccatccagga
     3241 agaccccagc acagactcac atatggacga ggatgggttt gagaaggacc ccttcccaaa
     3301 tagcagcaca gctgccaagt catttgagga tctcgcggac catccggtca ccagaagtga
     3361 aaaggctgcc tcctttaaac tgcagcgtca gaatcgtgtt gacagcaaag aaacagagtg
     3421 cggaggatct agagggccct acccatacga tgttccagat tacgcttaac tcgagctagc
     3481 ggccgcggct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     3541 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3601 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3661 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3721 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3781 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3841 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3901 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3961 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4021 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4081 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4141 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4201 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4321 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4381 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4441 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4501 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4561 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4621 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4681 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4741 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4801 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4861 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4921 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4981 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5041 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5101 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5161 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5221 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5281 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5341 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5401 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5461 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5521 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5581 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5641 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5701 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5761 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5821 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5881 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5941 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6001 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6061 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6121 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6181 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6241 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6301 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6361 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6421 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6481 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6541 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6601 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6661 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6721 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6781 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6841 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6901 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6961 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7021 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7081 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7141 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7201 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7261 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7321 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7381 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7441 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7501 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7561 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7621 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7681 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7741 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7801 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7861 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7921 gtgccacctg acgtc
//
