LOCUS       sequence-251333-        7138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human TACE (ADAM17).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7138)
  AUTHORS   Liu C, Xu P, Lamouille S, Xu J, Derynck R
  TITLE     TACE-mediated ectodomain shedding of the type I TGF-beta receptor 
            downregulates TGF-beta signaling.
  JOURNAL   Mol Cell. 2009 Jul 10. 35(1):26-36.
  PUBMED    19595713
REFERENCE   2  (bases 1 to 7138)
  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: "Mol Cell. 
            2009 Jul 10. 35(1):26-36."
COMMENT     Sequence Label: pRK5F-TACE
FEATURES             Location/Qualifiers
     source          1..7138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3403..3426
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3438..3447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3451..3585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3488..3507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3542..3561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3654..4011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3654..3845
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3677..3697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4031..4048)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4031..4047)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4040..4062)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4260..4715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4347..4366)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4557..4578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4732..4751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4851..4873
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4911..4929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4997..5101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5102..5170,5171..5962)
                     /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     complement(5320..5339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6133..6721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6622..6641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6875..6892
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7009..7030
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7045..7050,7051..7068,7069..7075)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7083..7099
                     /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)."
     primer_bind     7088..7110
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7107..7123
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7107..7123
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgatgaa catgaggcag tctctcctat tcctgaccag cgtggttcct
      961 ttcgtgctgg cgccgcgacc tccggatgac ccgggcttcg gcccccacca gagactcgag
     1021 aagcttgatt ctttgctctc agactacgat attctctctt tatctaatat ccagcagcat
     1081 tcggtaagaa aaagagatct acagacttca acacatgtag aaacactact aactttttca
     1141 gctttgaaaa ggcattttaa attatacctg acatcaagta ctgaacgttt ttcacaaaat
     1201 ttcaaggtcg tggtggtgga tggtaaaaac gaaagcgagt acactgtaaa atggcaggac
     1261 ttcttcactg gacacgtggt tggtgagcct gactctaggg ttctagccca cataagagat
     1321 gatgatgtta taatcagaat caacacagat ggggccgaat ataacataga gccactttgg
     1381 agatttgtta atgataccaa agacaaaaga atgttagttt ataaatctga agatatcaag
     1441 aatgtttcac gtttgcagtc tccaaaagtg tgtggttatt taaaagtgga taatgaagag
     1501 ttgctcccaa aagggttagt agacagagaa ccacctgaag agcttgttca tcgagtgaaa
     1561 agaagagctg acccagatcc catgaagaac acgtgtaaat tattggtggt agcagatcat
     1621 cgcttctaca gatacatggg cagaggggaa gagagtacaa ctacaaatta cttaatagag
     1681 ctaattgaca gagttgatga catctatcgg aacacttcat gggataatgc aggttttaaa
     1741 ggctatggaa tacagataga gcagattcgc attctcaagt ctccacaaga ggtaaaacct
     1801 ggtgaaaagc actacaacat ggcaaaaagt tacccaaatg aagaaaagga tgcttgggat
     1861 gtgaagatgt tgctagagca atttagcttt gatatagctg aggaagcatc taaagtttgc
     1921 ttggcacacc ttttcacata ccaagatttt gatatgggaa ctcttggatt agcttatgtt
     1981 ggctctccca gagcaaacag ccatggaggt gtttgtccaa aggcttatta tagcccagtt
     2041 gggaagaaaa atatctattt gaatagtggt ctgacgagca caaagaatta tggtaaaacc
     2101 atccttacaa aggaagctga cctggttaca actcatgaat tgggacataa ttttggagca
     2161 gaacatgatc cggatggtct agcagaatgt gccccgaatg aggaccaggg agggaaatat
     2221 gtcatgtatc ccatagctgt gagtggcgat cacgagaaca ataagatgtt ttcaaactgc
     2281 agtaaacaat caatctataa gaccattgaa agtaaggccc aggagtgttt tcaagaacgc
     2341 agcaataaag tttgtgggaa ctcgagggtg gatgaaggag aagagtgtga tcctggcatc
     2401 atgtatctga acaacgacac ctgctgcaac agcgactgca cgttgaagga aggtgtccag
     2461 tgcagtgaca ggaacagtcc ttgctgtaaa aactgtcagt ttgagactgc ccagaagaag
     2521 tgccaggagg cgattaatgc tacttgcaaa ggcgtgtcct actgcacagg taatagcagt
     2581 gagtgcccgc ctccaggaaa tgctgaagat gacactgttt gcttggatct tggcaagtgt
     2641 aaggatggga aatgcatccc tttctgcgag agggaacagc agctggagtc ctgtgcatgt
     2701 aatgaaactg acaactcctg caaggtgtgc tgcagggacc tttctggccg ctgtgtgccc
     2761 tatgtcgatg ctgaacaaaa gaacttattt ttgaggaaag gaaagccctg tacagtagga
     2821 ttttgtgaca tgaatggcaa atgtgagaaa cgagtacagg atgtaattga acgattttgg
     2881 gatttcattg accagctgag catcaatact tttggaaagt ttttagcaga caacatcgtt
     2941 gggtctgtcc tggttttctc cttgatattt tggattcctt tcagcattct tgtccattgt
     3001 gtggataaga aattggataa acagtatgaa tctctgtctc tgtttcaccc cagtaacgtc
     3061 gaaatgctga gcagcatgga ttctgcatcg gttcgcatta tcaaaccctt tcctgcgccc
     3121 cagactccag gccgcctgca gcctgcccct gtgatccctt cggcgccagc agctccaaaa
     3181 ctggaccacc agagaatgga caccatccag gaagacccca gcacagactc acatatggac
     3241 gaggatgggt ttgagaagga ccccttccca aatagcagca cagctgccaa gtcatttgag
     3301 gatctcacgg accatccggt caccagaagt gaaaaggctg cctcctttaa actgcagcgt
     3361 cagaatcgtg ttgacagcaa agaaacagag tgcgtcgaca tggactacaa ggacgacgac
     3421 gacaagtgat aagcttggcc gccatggccc aacttgttta ttgcagctta taatggttac
     3481 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3541 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcgggaa ttaattcggc
     3601 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3661 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctgtta
     4021 acagcttggc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4081 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4141 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     4201 attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta
     4261 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4321 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4381 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4441 tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc
     4501 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4561 actcttgttc caaactggaa caacactcaa ccctatctcg ggctattctt ttgatttata
     4621 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4681 cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg
     4741 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4801 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4861 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4921 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4981 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5041 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5101 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5161 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5221 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5281 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5341 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5401 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5461 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5521 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5581 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5641 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5701 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5761 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5821 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5881 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5941 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6001 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6061 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6121 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6181 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6241 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6301 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6361 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6421 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6481 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6541 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6601 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6661 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6721 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6781 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6841 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6901 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     6961 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     7021 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     7081 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaattaa
//
