LOCUS       sequence-384135-        7540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression ADAM10 in mammalian cells. Contains 
            metalloprotease, disintegrin, and cysteine rich domains--amino acids
            220-654.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7540)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 7540)
  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: pSCM167-Venus-VF-hADAM10(aa220-654)
FEATURES             Location/Qualifiers
     source          1..7540
                     /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"
     CDS             join(664..666,667..669,670..1380)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1420..1443
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    1465..1489
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2816..2840)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2859..2877)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2860..2877)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2897..2914)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2903..3127
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3173..3601
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3260..3279)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3470..3491
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3610..3630)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3615..3944
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3795..3930
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3857..3876
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4011..4805
                     /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(4065..4084)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4675..4694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4979..5100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5016..5035)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5070..5089
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5149..5165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5149..5165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5162..5184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5173..5189
                     /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(5197..5203,5204..5221,5222..5227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5242..5263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5380..5397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5551..6139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5631..5650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6310..7101,7102..7170))
                     /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     6933..6952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7171..7275)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7350..7369)
                     /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 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag
     1141 aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 ggagggggtg gatctggagg gggtggatct aagcttatgg actacaagga cgacgatgac
     1441 aagggtggtg gatccgaatt atcaacaagt ttgtacaaaa aagcaggctt cgaaggagat
     1501 agaaccatgg ccacttgtca gctttatatt cagactgatc atttgttctt taaatattac
     1561 ggaacacgag aagctgtgat tgcccagata tccagtcatg ttaaagcgat tgatacaatt
     1621 taccagacca cagacttctc cggaatccgt aacatcagtt tcatggtgaa acgcataaga
     1681 atcaatacaa ctgctgatga gaaggaccct acgaatcctt tccgtttccc aaatattggt
     1741 gtggagaagt ttctggaatt gaattctgag cagaatcatg atgactactg tttggcccat
     1801 gtcttcacag accgagattt tgatgatggc gtacttggtc tggcttgggt tggagcacct
     1861 tcaggaagct ctggaggaat atgtgaaaaa agtaaactct attcagatgg taagaagaag
     1921 tccttaaaca ctggaattat tactgttcag aactatgggt ctcatgtacc tcccaaagtc
     1981 tctcacatta cttttgctca cgaagttgga cataactttg gatccccaca tgattctgga
     2041 acagagtgca caccaggaga atctaagaat ttgggtcaaa aagaaaatgg caattacatc
     2101 atgtatgcaa gagcaacatc tggggacaaa cttaacaaca ataaattctc actctgtagt
     2161 attagaaata taagccaagt tcttgagaag aagagaaaca actgttttgt tgaatctggc
     2221 caacctattt gtggaaatgg aatggtagaa caaggtgaag aatgtgattg tggctatagt
     2281 gaccagtgta aagatgaatg ctgcttcgat gcaaatcaac cagagggaag aaaatgcaaa
     2341 ctgaaacctg ggaaacagtg cagtccaagt caaggtcctt gttgtacagc acagtgtgca
     2401 ttcaagtcaa agtctgagaa gtgtcgggat gattcagact gtgcaaggga aggaatatgt
     2461 aatggcttca cagctctctg cccagcatct gaccctaaac caaacttcac agactgtaat
     2521 aggcatacac aagtgtgcat taatgggcaa tgtgcaggtt ctatctgtga gaaatatggc
     2581 ttagaggagt gtacgtgtgc cagttctgat ggcaaagatg ataaagaatt atgccatgta
     2641 tgctgtatga agaaaatgga cccatcaact tgtgccagta cagggtctgt gcagtggagt
     2701 aggcacttca gtggtcgaac catcaccctg caacctggat ccccttgcaa cgattttaga
     2761 ggttactgtg atgtcttcat gcggtgcaga ttagtagatg ctgatggtcc tctaaaccca
     2821 gctttcttgt acaaagtggt tgatctagag ggccctattc tatagtgtca cctaaatgct
     2881 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2941 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3001 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3061 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3121 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3181 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3241 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3301 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3361 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3421 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3481 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3541 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3601 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3661 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3721 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3781 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3841 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3901 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3961 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4021 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4081 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4141 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4201 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4261 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4321 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4381 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4441 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4501 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4561 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4621 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4681 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4741 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4801 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4861 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4921 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4981 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5041 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5101 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5281 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5341 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     7321 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     7381 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     7441 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     7501 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
