LOCUS       sequence-349347-        4375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human SIGMAR1 at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4375)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4375)
  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: SIGMAR1_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(23..40)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(194..782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(274..293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(953..1744,1745..1813))
                     /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     1576..1595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1814..1918)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1986..2004
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2042..2064)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2164..2183
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2377..2399
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2391..2408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2392..2408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2860..3747
                     /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(4167..4183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4167..4183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4180..4202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4191..4207
                     /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(4215..4221,4222..4239,4240..4245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4260..4281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
       61 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      121 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
      421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
      481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
      541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
      601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
      661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
      721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
      781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
      841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
      901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
      961 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1021 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1081 caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1141 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1201 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1261 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1321 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1381 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     1441 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     1501 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     1561 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     1621 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     1681 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     1741 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     1801 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     1861 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1921 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1981 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     2041 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     2101 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     2161 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     2221 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     2281 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     2341 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     2401 cggccagtga attctgccac agaacattta ggagtggcca tctgggccag cccagtgagg
     2461 tagggctttt tgtttttctg aagctccctc tccattcccc acccctagtt agtccatggg
     2521 ccagggaagc agcagccatg gaagggcagg gcagagctgg cttttcactt gggtgctccc
     2581 tgtcccccag gggagacggt agtacacggg cctggtgagg caacagctgt ggagtggggg
     2641 ccaaacacat ggatggtgga gtacggccgg ggcgtcatcc catccaccct ggccttcgcg
     2701 ctggccgaca ctgtcttcag cacccaggac ttcctcaccc tcttctatac tcttcgctcc
     2761 tatgctcggg gcctccggct tgagctcacc acctacctct ttggccagga ccctggtgga
     2821 ggaggttctg gaggcggtgg aagtggtggc ggaggtagcg ccgagatcgg caccggcttt
     2881 cccttcgatc ctcactacgt ggaagtgctg ggcgagagaa tgcactatgt ggacgtgggc
     2941 cccagagatg gaacccctgt gctgtttctg cacggcaacc ctaccagcag ctacgtgtgg
     3001 cggaacatca tccctcacgt ggcccctaca cacagatgta tcgcccctga cctgatcggc
     3061 atgggcaaga gcgataagcc cgacctgggc tacttcttcg acgaccacgt gcggttcatg
     3121 gacgccttta ttgaggccct cggcctggaa gaggtggtgc tggttattca cgattggggc
     3181 agcgccctgg gattccactg ggccaagaga aaccccgaga gagtgaaggg aatcgccttc
     3241 atggaattca tcaggcccat tcctacctgg gacgagtggc ctgagttcgc cagagagaca
     3301 ttccaggcct tcagaaccac cgacgtgggc agaaagctga tcatcgacca gaacgtgttc
     3361 atcgagggca ccctgcctat gggagtcgtc agaccactga ccgaggtgga aatggaccac
     3421 tacagagagc cctttctgaa ccccgtggac cgggaacctc tttggagatt ccccaacgag
     3481 ctgcccattg ccggcgagcc tgctaatatt gtggccctgg tggaagagta catggactgg
     3541 ctgcatcaga gccccgtgcc taagctgctg ttttggggaa cacccggcgt gctgattcct
     3601 cctgctgaag ctgccagact ggccaagagc ctgcctaatt gcaaggccgt ggatatcggc
     3661 cctggcctga atctgctgca agaggacaac cccgatctga tcggctccga gattgccaga
     3721 tggctgagca ccctggaaat cagcggataa ccagccaggc ctgaaggaag acctgcggat
     3781 ggacaggagc gggcaggccc gcacatatcc acttgctgga gcccatgttt acagacaggg
     3841 acatacacca tgcagatcct gagttcctgc tgtatgagca gggatatcca tgcttatgta
     3901 tccaaacaca gagacccatg ggaacaaatg agacacatat agatactgag acctgtgtgt
     3961 acagtaggac catgcactca cacccatctg gagagggagc ccccggtata ccaagggagc
     4021 cagttgtgtt cagacacaca catcacagct tgactcacta actgaggcct ttccatagct
     4081 ccacagcttc ccacctcctc cccaccaaac cggggttcta gagttaagga tgggggaggg
     4141 tattatactg aagcttggcg taatctgtca tagctgtttc ctgtgtgaaa ttgttatccg
     4201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgc
//
