LOCUS       sequence-349365-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human CSF1R at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4442)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4442)
  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: CSF1R_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(543..1334,1335..1403))
                     /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     1166..1185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1404..1508)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1576..1594
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1632..1654)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1754..1773
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1967..1989
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1981..1998
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1982..1998
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2479..3366
                     /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(3824..3840)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3824..3840)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3837..3859)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3848..3864
                     /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(3872..3878,3879..3896,3897..3902))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3917..3938
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4055..4072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4226..4442,1..372))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4306..4325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
       61 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      361 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
      421 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
      481 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
      541 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
      601 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
      661 cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt atcagcaata
      721 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
      781 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
      841 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
      901 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
      961 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1021 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1081 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1141 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1201 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     1261 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     1321 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     1381 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     1441 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     1501 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     1561 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     1621 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     1681 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     1741 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     1801 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     1861 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     1921 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     1981 tgtaaaacga cggccagtga attcgagctc ggtacctcgc gaatgcatct agactaggga
     2041 gctgcccatg ggcatcctct gtcctatctc ccagatacag catcatgcag gcctgctggg
     2101 ccttggagcc cacccacaga cccaccttcc agcagatctg ctccttcctt caggagcagg
     2161 cccaagagga caggagagag cgggtgagtg gggtgaggct tggggtgggt ggccggtaaa
     2221 gcacgttggg ctgggcctga tggatctgga ctgacagttt ctggtccctc ccaccctcag
     2281 gactatacca atctgccgag cagcagcaga agcggtggca gcggcagcag cagcagtgag
     2341 ctggaggagg agagctctag tgagcacctg acctgctgcg agcaagggga tatcgcccag
     2401 cccttgctgc agcccaacaa ctatcagttc tgcggtggag gaggttctgg aggcggtgga
     2461 agtggtggcg gaggtagcgc cgagatcggc accggctttc ccttcgatcc tcactacgtg
     2521 gaagtgctgg gcgagagaat gcactatgtg gacgtgggcc ccagagatgg aacccctgtg
     2581 ctgtttctgc acggcaaccc taccagcagc tacgtgtggc ggaacatcat ccctcacgtg
     2641 gcccctacac acagatgtat cgcccctgac ctgatcggca tgggcaagag cgataagccc
     2701 gacctgggct acttcttcga cgaccacgtg cggttcatgg acgcctttat tgaggccctc
     2761 ggcctggaag aggtggtgct ggttattcac gattggggca gcgccctggg attccactgg
     2821 gccaagagaa accccgagag agtgaaggga atcgccttca tggaattcat caggcccatt
     2881 cctacctggg acgagtggcc tgagttcgcc agagagacat tccaggcctt cagaaccacc
     2941 gacgtgggca gaaagctgat catcgaccag aacgtgttca tcgagggcac cctgcctatg
     3001 ggagtcgtca gaccactgac cgaggtggaa atggaccact acagagagcc ctttctgaac
     3061 cccgtggacc gggaacctct ttggagattc cccaacgagc tgcccattgc cggcgagcct
     3121 gctaatattg tggccctggt ggaagagtac atggactggc tgcatcagag ccccgtgcct
     3181 aagctgctgt tttggggaac acccggcgtg ctgattcctc ctgctgaagc tgccagactg
     3241 gccaagagcc tgcctaattg caaggccgtg gatatcggcc ctggcctgaa tctgctgcaa
     3301 gaggacaacc ccgatctgat cggctccgag attgccagat ggctgagcac cctggaaatc
     3361 agcggataag gagttgacga cagggagtac cactctcccc tcccacaaac ttcaactcct
     3421 ccatggatgg ggcgacacgg ggagaacata caaactctgc cttcggtcat ttcactcaac
     3481 agctcggccc agctctgaaa cttgggaagg tgagggattc aggggaggtc agaggatccc
     3541 acttcctgag catgggccat cactgccagt caggggctgg gggctgagcc ctcacccccc
     3601 cctcccctac tgttctcatg gtgttggcct cgtgtttgct atgccaacta gtagaacctt
     3661 ctttcctaat ccccttatct tcatggaaat ggactgactt tatgcctatg aagtccccag
     3721 gagctacact gatactgaga aaaccaggct ctttggggct agacagactg gatcccgggc
     3781 ccgtcgactg cagaggcctg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
     3841 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3901 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3961 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     4021 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4441 tc
//
