LOCUS       sequence-384138-        8723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression ADAM10 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8723)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 8723)
  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"
COMMENT     Sequence Label: pDEST27-GST-hADAM10
FEATURES             Location/Qualifiers
     source          1..8723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        15..318
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        319..522
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     473..493
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     519..543
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             632..1285
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MAPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1243..1265
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     protein_bind    1313..1337
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3596..3620)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3652..3671)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3653..3671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3682..3699)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3682..3698)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3691..3713)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    4103..4237
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4140..4159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4194..4213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4363..4818
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4450..4469)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4660..4681
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4935..5292
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4935..5126
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4958..4978)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5143..5278
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5343..6137
                     /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(5397..5416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6007..6026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6201..6249
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6290..6309)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6409..6431
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        6555..6659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6660..6728,6729..7520)
                     /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(6878..6897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7691..8279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8180..8199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8433..8450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(8524..8628)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattagatgc atgtcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
       61 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      121 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      181 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      241 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      301 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      361 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      421 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      481 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
      541 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      601 agcctccgga ctctagccta ggccgcggac catggcccct atactaggtt attggaaaat
      661 taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa aatatgaaga
      721 gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg aattgggttt
      781 ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac agtctatggc
      841 catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa aagagcgtgc
      901 agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt cgagaattgc
      961 atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac ctgaaatgct
     1021 gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc atgtaaccca
     1081 tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc caatgtgcct
     1141 ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc cacaaattga
     1201 taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc aagccacgtt
     1261 tggtggtggc gaccatcctc caaaatcgga tctggttccg cgttctagat caacaagttt
     1321 gtacaaaaaa gcaggcttcg aaggagatag aaccatggcc gtgttgctga gagtgttaat
     1381 tctgctcctc tcctgggcgg cggggatggg aggtcagtat gggaatcctt taaataaata
     1441 tatcagacat tatgaaggat tatcttacaa tgtggattca ttacaccaaa aacaccagcg
     1501 tgccaaaaga gcagtctcac atgaagacca attttcacgt ctagatttcc atgcccatgg
     1561 aagacatttc aacctacgaa tgaagaggga cacttccctt ttcagtgatg aatttaaagt
     1621 agaaacatca aataaagtac ttggttatga tacctctcat atttacactg gacatattta
     1681 tggtgaagaa ggaagttttg gccatgggtc tgttattgat ggaagatttg aaggattcat
     1741 ccagactcgt ggtggcacat tttatgttga gccagcagag agatatatta aagaccgaac
     1801 tctgccattt cactctgtca tttatcatga agatgatatt aactatcccc ataaatacgg
     1861 tcctcagggg ggctgtgcag atcattcagt atttgaaaga atgaggaaat accagatgac
     1921 tggtgtagag gaagtaacac agatacctca agaagaacat gctgctaatg gtccagaact
     1981 tctgaggaaa aaacgtacaa cttcagctga aaaaaatact tgtcagcttt atattcagac
     2041 tgatcatttg ttctttaaat attacggaac acgagaagct gtgattgccc agatatccag
     2101 tcatgttaaa gcgattgata caatttacca gaccacagac ttctccggaa tccgtaacat
     2161 cagtttcatg gtgaaacgca taagaatcaa tacaactgct gatgagaagg accctacgaa
     2221 tcctttccgt ttcccaaata ttggtgtgga gaagtttctg gaattgaatt ctgagcagaa
     2281 tcatgatgac tactgtttgg cccatgtctt cacagaccga gattttgatg atggcgtact
     2341 tggtctggct tgggttggag caccttcagg aagctctgga ggaatatgtg aaaaaagtaa
     2401 actctattca gatggtaaga agaagtcctt aaacactgga attattactg ttcagaacta
     2461 tgggtctcat gtacctccca aagtctctca cattactttt gctcacgaag ttggacataa
     2521 ctttggatcc ccacatgatt ctggaacaga gtgcacacca ggagaatcta agaatttggg
     2581 tcaaaaagaa aatggcaatt acatcatgta tgcaagagca acatctgggg acaaacttaa
     2641 caacaataaa ttctcactct gtagtattag aaatataagc caagttcttg agaagaagag
     2701 aaacaactgt tttgttgaat ctggccaacc tatttgtgga aatggaatgg tagaacaagg
     2761 tgaagaatgt gattgtggct atagtgacca gtgtaaagat gaatgctgct tcgatgcaaa
     2821 tcaaccagag ggaagaaaat gcaaactgaa acctgggaaa cagtgcagtc caagtcaagg
     2881 tccttgttgt acagcacagt gtgcattcaa gtcaaagtct gagaagtgtc gggatgattc
     2941 agactgtgca agggaaggaa tatgtaatgg cttcacagct ctctgcccag catctgaccc
     3001 taaaccaaac ttcacagact gtaataggca tacacaagtg tgcattaatg ggcaatgtgc
     3061 aggttctatc tgtgagaaat atggcttaga ggagtgtacg tgtgccagtt ctgatggcaa
     3121 agatgataaa gaattatgcc atgtatgctg tatgaagaaa atggacccat caacttgtgc
     3181 cagtacaggg tctgtgcagt ggagtaggca cttcagtggt cgaaccatca ccctgcaacc
     3241 tggatcccct tgcaacgatt ttagaggtta ctgtgatgtc ttcatgcggt gcagattagt
     3301 agatgctgat ggtcctctag ctaggcttaa aaaagcaatt tttagtccag agctctatga
     3361 aaacattgct gaatggattg tggctcattg gtgggcagta ttacttatgg gaattgctct
     3421 gatcatgcta atggctggat ttattaagat atgcggtgtt catactccaa gtagtaatcc
     3481 aaagctgcct cctcctaaac cacttccagg cactttaaag aggaggagac ctccacagcc
     3541 cattcagcaa ccccagcgtc agcggccccg agagagttat caaatgggac acatgagacg
     3601 cgtttcttgt acaaagtggt tgatcgcgtg catgcgacgt catagctctc tccctatagt
     3661 gagtcgtatt ataagctagg cactggccgt cgttttacaa cgtcgtgact gggaaaactg
     3721 ctagcttggg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact
     3781 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa
     3841 actagctgca tatgcttgct gcttgagagt tttgcttact gagtatgatt tatgaaaata
     3901 ttatacacag gagctagtga ttctaattgt ttgtgtattt tagattcaca gtcccaaggc
     3961 tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc ataccacatt
     4021 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     4081 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     4141 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4201 gtccaaactc atcaatgtat cttatcatgt ctggatcgat cctgcattaa tgaatcggcc
     4261 aacgcgcggg gagaggcggt ttgcgtattg gctggcgtaa tagcgaagag gcccgcaccg
     4321 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4381 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4441 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4501 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4561 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4621 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4681 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4741 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4801 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4861 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     4921 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5101 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5161 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5221 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5281 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     5341 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5401 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     5461 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5521 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5581 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5641 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5701 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5761 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5821 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5881 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5941 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6001 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6061 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6121 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6181 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6241 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
     6301 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6361 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6421 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     6481 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     6541 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6601 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     6661 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     6721 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     6781 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     6841 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     6901 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     6961 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7021 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7081 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7141 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7201 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7261 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7321 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7381 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7441 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     7501 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     7561 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     7621 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7681 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7741 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7801 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     7861 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7921 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7981 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8041 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8101 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8161 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8221 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8281 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     8341 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     8401 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     8461 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagagctt
     8521 gcaattcgcg ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8581 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8641 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     8701 gcgtattacg aggccctttc acg
//
