LOCUS       sequence-349322-        4424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human SORL1 at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4424)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4424)
  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: SORL1_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(160..177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(331..919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(411..430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1090..1881,1882..1950))
                     /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     1713..1732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1951..2055)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2123..2141
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2179..2201)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2301..2320
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2514..2536
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2528..2545
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2529..2545
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3026..3913
                     /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(4353..4369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4353..4369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4366..4388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4377..4393
                     /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(4401..4407,4408..4424,1,2..7))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
       61 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      121 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      181 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      241 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      301 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      361 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      421 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      481 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      541 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      601 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      661 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      721 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      781 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      841 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      901 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      961 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1021 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1081 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1141 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     1201 atctggcccc agtgctgcaa tgataccgcg agatccacgc tcaccggctc cagatttatc
     1261 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     1321 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     1381 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     1441 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     1501 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     1561 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     1621 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     1681 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     1741 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     1801 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     1861 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     1921 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     1981 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     2041 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     2101 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     2161 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     2221 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     2281 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     2341 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2521 acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acctcgcgaa tgcatctaga
     2581 tatagataac tcaaactggg gaacccaaac tacccactta gcttttgagt aggtaataag
     2641 agtcttctcc tgagaagctt tgttttccta atttggctac gttcaaacga ccactaaaat
     2701 aaatgtgagg atccatcgat gggtagaagg tggccaggag ggacagtggg aagctgggct
     2761 aggtttcaca gctgattatg gtggctgcct ggctatggca ttgaccttct cagctagagg
     2821 gttgtggtag ggttgagggg tgggtagagt ggtgggatat ggggtcaggt gaaaatgtgt
     2881 tggaactcac caaggcctga ctgttttgtc tctaggggaa gatgatgaag atgcccctat
     2941 gataactgga ttttcagatg acgtccccat ggtgatagcc ggtggaggag gttctggagg
     3001 cggtggaagt ggtggcggag gtagcgccga gatcggcacc ggctttccct tcgatcctca
     3061 ctacgtggaa gtgctgggcg agagaatgca ctatgtggac gtgggcccca gagatggaac
     3121 ccctgtgctg tttctgcacg gcaaccctac cagcagctac gtgtggcgga acatcatccc
     3181 tcacgtggcc cctacacaca gatgtatcgc ccctgacctg atcggcatgg gcaagagcga
     3241 taagcccgac ctgggctact tcttcgacga ccacgtgcgg ttcatggacg cctttattga
     3301 ggccctcggc ctggaagagg tggtgctggt tattcacgat tggggcagcg ccctgggatt
     3361 ccactgggcc aagagaaacc ccgagagagt gaagggaatc gccttcatgg aattcatcag
     3421 gcccattcct acctgggacg agtggcctga gttcgccaga gagacattcc aggccttcag
     3481 aaccaccgac gtgggcagaa agctgatcat cgaccagaac gtgttcatcg agggcaccct
     3541 gcctatggga gtcgtcagac cactgaccga ggtggaaatg gaccactaca gagagccctt
     3601 tctgaacccc gtggaccggg aacctctttg gagattcccc aacgagctgc ccattgccgg
     3661 cgagcctgct aatattgtgg ccctggtgga agagtacatg gactggctgc atcagagccc
     3721 cgtgcctaag ctgctgtttt ggggaacacc cggcgtgctg attcctcctg ctgaagctgc
     3781 cagactggcc aagagcctgc ctaattgcaa ggccgtggat atcggccctg gcctgaatct
     3841 gctgcaagag gacaaccccg atctgatcgg ctccgagatt gccagatggc tgagcaccct
     3901 ggaaatcagc ggataaaaga gctttcctca ctagaaacca aatggtgtaa atattttatt
     3961 tgataaagat agttgatggt ttattttaaa agatgcactt tgagttgcaa tatgttattt
     4021 ttatatgggc caaaaacaaa aaacaaaaaa aaaaaaaagg aaagaaagga atgaataaac
     4081 tttgtagtaa tcaactgtga acttcaaacc aggttgattt tagtaaccca attgctttga
     4141 tttgacatta atgtagtctt acagggctgt gcttgctggg catgctttta cgtctgtgag
     4201 ataatttcgg ttcagtaaat tggccaatct ttttattttt ctaagacaca gaaatgtatt
     4261 taataaaaac ctcgagagag tgatgggtgg aaccccttct ccttgaaagt gtgtacctgc
     4321 agaggcctgc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4381 tatccgctca caattccaca caacatacga gccggaagca taaa
//
