LOCUS       sequence-349344-        4498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human ITM2B at the C-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4498)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4498)
  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: ITM2B_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(170..758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(250..269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(929..1720,1721..1789))
                     /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     1552..1571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1790..1894)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1962..1980
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2018..2040)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2140..2159
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2353..2375
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2368..2384
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2424..2440
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2892..3779
                     /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(4266..4282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4266..4282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4279..4301)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4290..4306
                     /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(4314..4320,4321..4338,4339..4344))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4359..4380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(4497..4498,1..16))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
       61 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
      421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct
     1081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     1561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     1921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     1981 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2041 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2101 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2161 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2221 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     2281 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     2341 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
     2401 cctcgcgaat gcatctagat gttgtaaaac gacggccagt gaattcttcc acaaagatac
     2461 acaaggtctt gcaagcacca ccatagtcaa gatacagaac attttcaata ccttcaagtt
     2521 catgaatttg taagcagtct ggatagcatt attaatgttg ctgtgatctg tcaagatatt
     2581 aatacttggt tcagaaaagg gagaaaaatt acttttcttc agtgtaaaca ttcaaactgt
     2641 agaagttttt gtggaatata gagtgaaata cttctatatg tctaaataat aactataaaa
     2701 atgatgtgaa tatttattat taaagaatca aaattttaaa aaactttttt ccctctccaa
     2761 caggtattca gaaacgtgaa gccagcaatt gtttcgcaat tcggcatttt gaaaacaaat
     2821 ttgccgtgga aactttaatt tgttctggtg gaggaggttc tggaggcggt ggaagtggtg
     2881 gcggaggtag cgccgagatc ggcaccggct ttcccttcga tcctcactac gtggaagtgc
     2941 tgggcgagag aatgcactat gtggacgtgg gccccagaga tggaacccct gtgctgtttc
     3001 tgcacggcaa ccctaccagc agctacgtgt ggcggaacat catccctcac gtggccccta
     3061 cacacagatg tatcgcccct gacctgatcg gcatgggcaa gagcgataag cccgacctgg
     3121 gctacttctt cgacgaccac gtgcggttca tggacgcctt tattgaggcc ctcggcctgg
     3181 aagaggtggt gctggttatt cacgattggg gcagcgccct gggattccac tgggccaaga
     3241 gaaaccccga gagagtgaag ggaatcgcct tcatggaatt catcaggccc attcctacct
     3301 gggacgagtg gcctgagttc gccagagaga cattccaggc cttcagaacc accgacgtgg
     3361 gcagaaagct gatcatcgac cagaacgtgt tcatcgaggg caccctgcct atgggagtcg
     3421 tcagaccact gaccgaggtg gaaatggacc actacagaga gccctttctg aaccccgtgg
     3481 accgggaacc tctttggaga ttccccaacg agctgcccat tgccggcgag cctgctaata
     3541 ttgtggccct ggtggaagag tacatggact ggctgcatca gagccccgtg cctaagctgc
     3601 tgttttgggg aacacccggc gtgctgattc ctcctgctga agctgccaga ctggccaaga
     3661 gcctgcctaa ttgcaaggcc gtggatatcg gccctggcct gaatctgctg caagaggaca
     3721 accccgatct gatcggctcc gagattgcca gatggctgag caccctggaa atcagcggat
     3781 aaacagtcaa gaaaaacatt attgaggaaa attaatatca cagcataacc gcacccttta
     3841 cattttgtgc agtgattatt ttttaaagtc ttctttcatg taagtagcaa acagggcttt
     3901 actatctttt catctcatta attcaattaa aaccattacc ttaaaatttt tttctttcga
     3961 agtgtggtgt cttttatatt tgaattagta actgtatgaa gtcatagata atagtacatg
     4021 tcaccttagg tagtaggaag aattacaatt tctttaaatc atttatctgg atttttatgt
     4081 tttattagca ttttcaagaa gacggattat ctagagaata atcatatata tgcatacgta
     4141 aaaatggacc acagtgactt atttgtagtt gttagttgcc ctaagcttgg cgtaatcatg
     4201 gtcatagcat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa
     4261 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4321 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4381 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcct
//
