LOCUS       sequence-349340-        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human ADAM10 at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4456)
  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: ADAM10_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(116..133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(287..875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(367..386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1046..1837,1838..1906))
                     /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     1669..1688
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1907..2011)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2079..2097
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2135..2157)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2257..2276
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2470..2492
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2484..2501
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2485..2501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2988..3875
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     primer_bind     complement(4341..4357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4341..4357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4354..4376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4365..4381
                     /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(4389..4395,4396..4413,4414..4419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4434..4455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
       61 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
      121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
      181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
      241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
      301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
      361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
      421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
      481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
      541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
      601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
      661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
      721 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
      781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
      901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
      961 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1021 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1081 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     1141 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat ccacgctcac
     1201 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     1261 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     1321 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     1381 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     1441 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     1501 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     1561 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     1621 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     1681 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     1741 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     1801 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     1861 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     1921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     1981 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     2041 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     2101 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     2161 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     2221 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     2281 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     2341 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     2401 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
     2461 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtacct
     2521 cgcgaatgca tctagatcca atcataccag actttgaaga tttagtataa aaaaagaaat
     2581 attttgttag taatttttgt attgattata tattgaaatg ataacacttt ggctatatta
     2641 atttaaataa aataaaaaat attattaaaa ttatttcacc ttttaaacct tttttattgt
     2701 ggctatcaga agattttaaa ttacattgat ggcccatatt attggacagc actgaactaa
     2761 aacctttctt attgaaacag cagctttgct tttaaagatt aaaaagctca tgataaataa
     2821 atctgctttg cttttatttg tggctcacag gcactttaaa gaggaggaga cctccacagc
     2881 ccattcagca accccagcgt cagcggcccc gagagagtta tcaaatggga cacatgagac
     2941 gcggtggagg aggttctgga ggcggtggaa gtggtggcgg aggtagcgcc gagatcggca
     3001 ccggctttcc cttcgatcct cactacgtgg aagtgctggg cgagagaatg cactatgtgg
     3061 acgtgggccc cagagatgga acccctgtgc tgtttctgca cggcaaccct accagcagct
     3121 acgtgtggcg gaacatcatc cctcacgtgg cccctacaca cagatgtatc gcccctgacc
     3181 tgatcggcat gggcaagagc gataagcccg acctgggcta cttcttcgac gaccacgtgc
     3241 ggttcatgga cgcctttatt gaggccctcg gcctggaaga ggtggtgctg gttattcacg
     3301 attggggcag cgccctggga ttccactggg ccaagagaaa ccccgagaga gtgaagggaa
     3361 tcgccttcat ggaattcatc aggcccattc ctacctggga cgagtggcct gagttcgcca
     3421 gagagacatt ccaggccttc agaaccaccg acgtgggcag aaagctgatc atcgaccaga
     3481 acgtgttcat cgagggcacc ctgcctatgg gagtcgtcag accactgacc gaggtggaaa
     3541 tggaccacta cagagagccc tttctgaacc ccgtggaccg ggaacctctt tggagattcc
     3601 ccaacgagct gcccattgcc ggcgagcctg ctaatattgt ggccctggtg gaagagtaca
     3661 tggactggct gcatcagagc cccgtgccta agctgctgtt ttggggaaca cccggcgtgc
     3721 tgattcctcc tgctgaagct gccagactgg ccaagagcct gcctaattgc aaggccgtgg
     3781 atatcggccc tggcctgaat ctgctgcaag aggacaaccc cgatctgatc ggctccgaga
     3841 ttgccagatg gctgagcacc ctggaaatca gcggataact gcagcttttg cgttggttct
     3901 tcctagtgcc tacaatggga aaacttcact ccaaagagaa acctattaag tcatcatctc
     3961 caaactaaac cctcacaagt aacagttgaa gaaaaaatgg caagagatca tatcctcaga
     4021 ccaggtggaa ttacttaaat tttaaagcct gaaaattcca atttgggggt gggaggtgga
     4081 aaaggaaccc aattttctta tgaacagata tttttaactt aatggcacaa agtcttagaa
     4141 tattattatg tgccccgtgt tccctgttct tcgttgctgc attttcttca cttgcaggca
     4201 aacttggctc tcaataaact tttaccacaa attgaaataa atatattttt ttcaactgcc
     4261 aatcaaggct aggaggctcc aatatcggat cccgggcccg tcgactgcag aggcctgcat
     4321 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4381 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4441 agctaactca cattaa
//
