LOCUS       sequence-349329-        4377 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human GBA at the C-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4377)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4377)
  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: GBA_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(9..15,16..33,34..39))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    54..75
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(192..209)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(363..951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(443..462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1122..1913,1914..1982))
                     /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     1745..1764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1983..2087)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2155..2173
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2211..2233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2333..2352
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2546..2568
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2560..2577
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2561..2577
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3029..3916
                     /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(4338..4354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4338..4354)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4351..4373)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    join(4362..4377,1)
                     /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)."
ORIGIN
        1 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
       61 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
      121 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      781 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1261 cgagatccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2161 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2341 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2401 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2461 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2521 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2581 ttcggtgggt ccgtgggtgg gtgacttctt agatgagggt ttcatgggag gtaccccgag
     2641 ggactctgac catctgttcc cacattcagc aagttcattc ctgagggctc ccagagagtg
     2701 gggctggttg ccagtcagaa gaacgacctg gacgcagtgg cactgatgca tcccgatggc
     2761 tctgctgttg tggtcgtgct aaaccggtga gggcaatggt gaggtctggg aagtgggctg
     2821 aagacagcgt tgggggcctt ggcaggatca cactctcagc ttctcctccc tgctccctag
     2881 ctcctctaag gatgtgcctc ttaccatcaa ggatcctgct gtgggcttcc tggagacaat
     2941 ctcacctggc tactccattc acacctacct gtggcgtcgc cagggtggag gaggttctgg
     3001 aggcggtgga agtggtggcg gaggtagcgc cgagatcggc accggctttc ccttcgatcc
     3061 tcactacgtg gaagtgctgg gcgagagaat gcactatgtg gacgtgggcc ccagagatgg
     3121 aacccctgtg ctgtttctgc acggcaaccc taccagcagc tacgtgtggc ggaacatcat
     3181 ccctcacgtg gcccctacac acagatgtat cgcccctgac ctgatcggca tgggcaagag
     3241 cgataagccc gacctgggct acttcttcga cgaccacgtg cggttcatgg acgcctttat
     3301 tgaggccctc ggcctggaag aggtggtgct ggttattcac gattggggca gcgccctggg
     3361 attccactgg gccaagagaa accccgagag agtgaaggga atcgccttca tggaattcat
     3421 caggcccatt cctacctggg acgagtggcc tgagttcgcc agagagacat tccaggcctt
     3481 cagaaccacc gacgtgggca gaaagctgat catcgaccag aacgtgttca tcgagggcac
     3541 cctgcctatg ggagtcgtca gaccactgac cgaggtggaa atggaccact acagagagcc
     3601 ctttctgaac cccgtggacc gggaacctct ttggagattc cccaacgagc tgcccattgc
     3661 cggcgagcct gctaatattg tggccctggt ggaagagtac atggactggc tgcatcagag
     3721 ccccgtgcct aagctgctgt tttggggaac acccggcgtg ctgattcctc ctgctgaagc
     3781 tgccagactg gccaagagcc tgcctaattg caaggccgtg gatatcggcc ctggcctgaa
     3841 tctgctgcaa gaggacaacc ccgatctgat cggctccgag attgccagat ggctgagcac
     3901 cctggaaatc agcggataat ggagcagata ctcaaggagg cactgggctc agcctgggca
     3961 ttaaagggac agagtcagct cacacgctgt ctgtgactaa agagggcaca gcagggccag
     4021 tgtgagctta cagcgacgta agcccagggg caatggtttg ggtgactcac tttcccctct
     4081 aggtggtgcc aggggctgga ggcccctaga aaaagatcag taagccccag tgtcccccca
     4141 gcccccatgc ttatgtgaac atgcgctgtg tgctgcttgc tttggaaact gggcctgggt
     4201 ccaggcctag ggtgagctca ctgtccgtac aaacacaaga tcagggctga gggtaaggaa
     4261 aagaagagac taggaaagct gggcccaaaa ctggagactg tttgtctttc ctggagatga
     4321 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcaca
//
