LOCUS       sequence-349082-        4439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human TIA1 at the N-terminus 
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4439)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4439)
  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: TIA1_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4439
                     /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             2844..3734
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(4231..4247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4231..4247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4244..4266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4255..4271
                     /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(4279..4285,4286..4303,4304..4309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4324..4345
                     /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 attcgagctc ggtacctcgc gaatgcatct agaaagttca cgcaaatcgt
     2461 gaagtagtta atccttgcgc ctgcgcagtc tgaggcgtcc ctcgcgtcgg gtggaagagc
     2521 ggctgcgcaa caagtgaacc tatggattaa ggtgtagccg ggcggcccgg aggttgagag
     2581 tgcaagatta ttctgcgcct gcgcggcccg ggtaactttg ctggctgctc ctaggctccc
     2641 ggctcgccgc catcttgtat tggggtttca ttgttcccgc tgggccgggc ggtttagtgt
     2701 aattgccgcc ggaggaggag gcggagtaac ctctggtcag ccgagaaacc ccactatcct
     2761 gtagccataa ccgcttaaac gatttgggag gtagtgaagg gcagggagct ggacctggag
     2821 gcgccgccgc gacagcagca gccatggccg agatcggcac cggctttccc ttcgatcctc
     2881 actacgtgga agtgctgggc gagagaatgc actatgtgga cgtgggcccc agagatggaa
     2941 cccctgtgct gtttctgcac ggcaacccta ccagcagcta cgtgtggcgg aacatcatcc
     3001 ctcacgtggc ccctacacac agatgtatcg cccctgacct gatcggcatg ggcaagagcg
     3061 ataagcccga cctgggctac ttcttcgacg accacgtgcg gttcatggac gcctttattg
     3121 aggccctcgg cctggaagag gtggtgctgg ttattcacga ttggggcagc gccctgggat
     3181 tccactgggc caagagaaac cccgagagag tgaagggaat cgccttcatg gaattcatca
     3241 ggcccattcc tacctgggac gagtggcctg agttcgccag agagacattc caggccttca
     3301 gaaccaccga cgtgggcaga aagctgatca tcgaccagaa cgtgttcatc gagggcaccc
     3361 tgcctatggg agtcgtcaga ccactgaccg aggtggaaat ggaccactac agagagccct
     3421 ttctgaaccc cgtggaccgg gaacctcttt ggagattccc caacgagctg cccattgccg
     3481 gcgagcctgc taatattgtg gccctggtgg aagagtacat ggactggctg catcagagcc
     3541 ccgtgcctaa gctgctgttt tggggaacac ccggcgtgct gattcctcct gctgaagctg
     3601 ccagactggc caagagcctg cctaattgca aggccgtgga tatcggccct ggcctgaatc
     3661 tgctgcaaga ggacaacccc gatctgatcg gctccgagat tgccagatgg ctgagcaccc
     3721 tggaaatcag cggaggagga ggttctggag gcggtggaag tggtggcgga ggtagcgagg
     3781 acgagatgcc caagactctg tgagtctggg gcagcgatga gggaggcggg atggtggtcg
     3841 tcccggaggg aaggcctcgg ccctgcgccg ctcccagcct attgttctcc gtggacaccg
     3901 cgatatcgtg gtttttgttt ccttagcatg cacctggatg cttacaacca agacccaatc
     3961 tgtgaggacg aggtgaaggg agacggggct caaccctctc aattaatccc ttcactttag
     4021 caacaccatt gcatgacctt tgctctcgcc acgggccctc tgtggtctta gacctggaga
     4081 aaagacttta attcagacct ctttttagaa ctcgtcgaac cttccttttg tcctatgctt
     4141 tctttccttt tcccgcttct cccccttttt gattaaggat cccgggcccg tcgactgcag
     4201 aggcctgcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4261 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4321 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4381 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgc
//
