LOCUS       sequence-374396-        7612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression ADAM10 in mammalian cells. Contains prodomain and
            metalloprotease domain--amino acids 1-459.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7612)
  TITLE     TREAT-AD plasmids 2022
REFERENCE   2  (bases 1 to 7612)
  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(aa1-459)
FEATURES             Location/Qualifiers
     source          1..7612
                     /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(2888..2912)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2931..2949)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2932..2949)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2969..2986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2975..3199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3245..3673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3332..3351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3542..3563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3682..3702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3687..4016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3867..4002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3929..3948
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4083..4877
                     /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(4137..4156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4747..4766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5051..5172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5088..5107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5142..5161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5221..5237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5221..5237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5234..5256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5245..5261
                     /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(5269..5275,5276..5293,5294..5299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5314..5335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5452..5469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5623..6211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5703..5722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6382..7173,7174..7242))
                     /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     7005..7024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7243..7347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7422..7441)
                     /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 ccgtgttgct gagagtgtta attctgctcc tctcctgggc ggcggggatg
     1561 ggaggtcagt atgggaatcc tttaaataaa tatatcagac attatgaagg attatcttac
     1621 aatgtggatt cattacacca aaaacaccag cgtgccaaaa gagcagtctc acatgaagac
     1681 caattttcac gtctagattt ccatgcccat ggaagacatt tcaacctacg aatgaagagg
     1741 gacacttccc ttttcagtga tgaatttaaa gtagaaacat caaataaagt acttggttat
     1801 gatacctctc atatttacac tggacatatt tatggtgaag aaggaagttt tggccatggg
     1861 tctgttattg atggaagatt tgaaggattc atccagactc gtggtggcac attttatgtt
     1921 gagccagcag agagatatat taaagaccga actctgccat ttcactctgt catttatcat
     1981 gaagatgata ttaactatcc ccataaatac ggtcctcagg ggggctgtgc agatcattca
     2041 gtatttgaaa gaatgaggaa ataccagatg actggtgtag aggaagtaac acagatacct
     2101 caagaagaac atgctgctaa tggtccagaa cttctgagga aaaaacgtac aacttcagct
     2161 gaaaaaaata cttgtcagct ttatattcag actgatcatt tgttctttaa atattacgga
     2221 acacgagaag ctgtgattgc ccagatatcc agtcatgtta aagcgattga tacaatttac
     2281 cagaccacag acttctccgg aatccgtaac atcagtttca tggtgaaacg cataagaatc
     2341 aatacaactg ctgatgagaa ggaccctacg aatcctttcc gtttcccaaa tattggtgtg
     2401 gagaagtttc tggaattgaa ttctgagcag aatcatgatg actactgttt ggcccatgtc
     2461 ttcacagacc gagattttga tgatggcgta cttggtctgg cttgggttgg agcaccttca
     2521 ggaagctctg gaggaatatg tgaaaaaagt aaactctatt cagatggtaa gaagaagtcc
     2581 ttaaacactg gaattattac tgttcagaac tatgggtctc atgtacctcc caaagtctct
     2641 cacattactt ttgctcacga agttggacat aactttggat ccccacatga ttctggaaca
     2701 gagtgcacac caggagaatc taagaatttg ggtcaaaaag aaaatggcaa ttacatcatg
     2761 tatgcaagag caacatctgg ggacaaactt aacaacaata aattctcact ctgtagtatt
     2821 agaaatataa gccaagttct tgagaagaag agaaacaact gttttgttga atctggccaa
     2881 cctattaacc cagctttctt gtacaaagtg gttgatctag agggccctat tctatagtgt
     2941 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3001 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3061 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3121 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3181 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3241 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3301 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3361 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3421 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3481 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3541 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3601 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3661 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3721 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3901 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3961 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4021 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4381 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4861 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4921 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4981 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5041 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5101 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5161 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5221 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5281 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5341 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5401 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5461 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5521 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5581 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5641 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5701 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5761 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5821 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5881 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5941 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6001 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6061 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6121 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6181 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6241 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6301 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6361 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6421 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6481 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6541 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6601 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6661 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6721 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6781 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6841 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6901 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6961 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7021 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7081 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7141 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7201 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7261 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7321 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7381 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7441 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7501 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7561 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
