LOCUS       sequence-349379-        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human CHMP2B at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4456)
  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: CHMP2B_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(376..393)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(376..392)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(385..407)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(601..620)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     720..742
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(780..798)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        866..970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(971..1039,1040..1831)
                     /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     complement(1189..1208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2002..2590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2491..2510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2744..2761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2878..2899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2914..2919,2920..2937,2938..2944)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2952..2968
                     /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)."
     primer_bind     2957..2979
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2976..2992
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2976..2992
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3500..4387
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
ORIGIN
        1 tcttttacaa aacacatgta ttttgcaaaa aaaaaaaaaa tgaagaccat gagtgaacag
       61 ttgtttccta acccatggct atttagaatc ttttgccaaa gaatgacaat gatgcaaaaa
      121 tgggaacagt ttggatttta attagaactg tttaggagtg atgatgtgta aaaagttgac
      181 ttctcttttg catggcacag agaaattata ttccttactt catgtcagtt tatgttctaa
      241 atctttttca ctgaatataa aaatcttgtt aaatgccatt aggcaccaac ttaaagaggg
      301 ttgtaaaaat attaaaagta tatcgttaat tctgccaata tctagatgca ttcgcgaggt
      361 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      421 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      481 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
      541 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
      601 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
      661 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
      721 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
      781 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1201 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1501 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1681 gcgtggatct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1861 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1921 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1981 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2041 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2101 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2161 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2221 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2281 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2341 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2401 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2461 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2521 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2581 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2641 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2701 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2761 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2821 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     2881 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     2941 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     3001 cgccaagctt gcatgcaggc ctctgcagtc gacgggcccg ggatccgatc caattgacat
     3061 ctttgacggt tctgatgacg aagaagaaag ccaggatatt gtgaatcaag ttcttgatga
     3121 aattggaatt gaaatttctg gaaaggtatg aacatcatct tttcttagtt ggaaatagtt
     3181 tctgcctacc acgtttgtca cttaattgtt ttgtttttac taggaggtgc atggttttta
     3241 tttctgtttc aaaagtaaat taaacagacc tctttacagc acatcctgta tgtcctagtc
     3301 caaatgtttc ctaatgcacg tttgtctttt tcattgttta atatagatgg ccaaagctcc
     3361 atcagctgct cgaagcttac catctgcctc tacttcaaag gctacaatct cagatgaaga
     3421 gattgaacgg caactcaagg ctttaggagt agatggtgga ggaggttctg gaggcggtgg
     3481 aagtggtggc ggaggtagcg ccgagatcgg caccggcttt cccttcgatc ctcactacgt
     3541 ggaagtgctg ggcgagagaa tgcactatgt ggacgtgggc cccagagatg gaacccctgt
     3601 gctgtttctg cacggcaacc ctaccagcag ctacgtgtgg cggaacatca tccctcacgt
     3661 ggcccctaca cacagatgta tcgcccctga cctgatcggc atgggcaaga gcgataagcc
     3721 cgacctgggc tacttcttcg acgaccacgt gcggttcatg gacgccttta ttgaggccct
     3781 cggcctggaa gaggtggtgc tggttattca cgattggggc agcgccctgg gattccactg
     3841 ggccaagaga aaccccgaga gagtgaaggg aatcgccttc atggaattca tcaggcccat
     3901 tcctacctgg gacgagtggc ctgagttcgc cagagagaca ttccaggcct tcagaaccac
     3961 cgacgtgggc agaaagctga tcatcgacca gaacgtgttc atcgagggca ccctgcctat
     4021 gggagtcgtc agaccactga ccgaggtgga aatggaccac tacagagagc cctttctgaa
     4081 ccccgtggac cgggaacctc tttggagatt ccccaacgag ctgcccattg ccggcgagcc
     4141 tgctaatatt gtggccctgg tggaagagta catggactgg ctgcatcaga gccccgtgcc
     4201 taagctgctg ttttggggaa cacccggcgt gctgattcct cctgctgaag ctgccagact
     4261 ggccaagagc ctgcctaatt gcaaggccgt ggatatcggc cctggcctga atctgctgca
     4321 agaggacaac cccgatctga tcggctccga gattgccaga tggctgagca ccctggaaat
     4381 cagcggataa tcaaaagaag tcatactatt ttgcttactt ataattatgt agtataaact
     4441 aagcacagtg cagatt
//
