LOCUS       sequence-349338-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human RAB 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: RAB_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1988..2006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2044..2066)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2166..2185
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2379..2401
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2393..2410
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2394..2410
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2891..3778
                     /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(4236..4252)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4236..4252)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4249..4271)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4260..4276
                     /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(4284..4290,4291..4308,4309..4314))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4329..4350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagatc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1981 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2041 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2101 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2161 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2221 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2281 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2341 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2401 gacggccagt gaattcgagc tcggtacctc gcgaatgcat ctagagaact cagtaggtgg
     2461 tcttctctta tttgacatta ccaaccgcag gtccttccag aatgtccatg agtggttaga
     2521 agagaccaaa gtacacgttc agccctacca aattgtattt gttctggtgg gtcacaagtg
     2581 tgacctggat acacagaggc aagtgactcg ccacgaggcc gagaaactgg ctgctgcata
     2641 cggcatgaag tacattgaaa cgtcagcccg agatgccatt aatgtggaga aagccttcac
     2701 agacctgaca agagacatat atgagctggt taaaaggggg gagattacaa tccaggaggg
     2761 ctgggaaggg gtgaagagtg gatttgtacc aaatgtggtt cactcttcag aagaggttgt
     2821 caaatcagag aggagatgtt tgtgcggtgg aggaggttct ggaggcggtg gaagtggtgg
     2881 cggaggtagc gccgagatcg gcaccggctt tcccttcgat cctcactacg tggaagtgct
     2941 gggcgagaga atgcactatg tggacgtggg ccccagagat ggaacccctg tgctgtttct
     3001 gcacggcaac cctaccagca gctacgtgtg gcggaacatc atccctcacg tggcccctac
     3061 acacagatgt atcgcccctg acctgatcgg catgggcaag agcgataagc ccgacctggg
     3121 ctacttcttc gacgaccacg tgcggttcat ggacgccttt attgaggccc tcggcctgga
     3181 agaggtggtg ctggttattc acgattgggg cagcgccctg ggattccact gggccaagag
     3241 aaaccccgag agagtgaagg gaatcgcctt catggaattc atcaggccca ttcctacctg
     3301 ggacgagtgg cctgagttcg ccagagagac attccaggcc ttcagaacca ccgacgtggg
     3361 cagaaagctg atcatcgacc agaacgtgtt catcgagggc accctgccta tgggagtcgt
     3421 cagaccactg accgaggtgg aaatggacca ctacagagag ccctttctga accccgtgga
     3481 ccgggaacct ctttggagat tccccaacga gctgcccatt gccggcgagc ctgctaatat
     3541 tgtggccctg gtggaagagt acatggactg gctgcatcag agccccgtgc ctaagctgct
     3601 gttttgggga acacccggcg tgctgattcc tcctgctgaa gctgccagac tggccaagag
     3661 cctgcctaat tgcaaggccg tggatatcgg ccctggcctg aatctgctgc aagaggacaa
     3721 ccccgatctg atcggctccg agattgccag atggctgagc accctggaaa tcagcggata
     3781 atcagttctt ttatttccaa aacatgctct cctacttgaa ctgaaaagta agagaaataa
     3841 atagaatctt tgtgtaactg atgagaactc aacctccatt tggataccaa gtgagcctcc
     3901 ctctcagggg gactctgaca gactcgtgtg cgggtggatg ggggtgcctg agtgctgtga
     3961 ccaagggact gaattgctga catgggccag gccaggcact gttttgtggc ccatgctagg
     4021 cagtggctgc tccatgagcc ctttgtggac cacatgttaa aggcctgggg tttaggaagc
     4081 taagtcccag gaaaacatac ctacagacag gacaactgtt ccttggtatt tgagctcact
     4141 tataggccat gaacataaag aaatacataa aggggaaaaa aggatcccgg gcccgtcgac
     4201 tgcagaggcc tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4261 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4321 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4381 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4441 tt
//
