LOCUS       sequence-349368-        4442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for endogenous tagging of human OPTN 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: OPTN_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             169..1056
                     /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(1514..1530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1514..1530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1527..1549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1538..1554
                     /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(1562..1568,1569..1586,1587..1592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1607..1628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1745..1762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1916..2504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1996..2015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2675..3466,3467..3535))
                     /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     3298..3317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3536..3640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3708..3726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3764..3786)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3886..3905
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4099..4121
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4113..4130
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4114..4130
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccaggagctg aggacaggga ctggcggcaa cagcggaata ttccgattca ttcctgcccc
       61 aagtgtggag aggttctgcc tgacatagac acgttacaga ttcacgtgat ggattgcatc
      121 attggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcgc cgagatcggc
      181 accggctttc ccttcgatcc tcactacgtg gaagtgctgg gcgagagaat gcactatgtg
      241 gacgtgggcc ccagagatgg aacccctgtg ctgtttctgc acggcaaccc taccagcagc
      301 tacgtgtggc ggaacatcat ccctcacgtg gcccctacac acagatgtat cgcccctgac
      361 ctgatcggca tgggcaagag cgataagccc gacctgggct acttcttcga cgaccacgtg
      421 cggttcatgg acgcctttat tgaggccctc ggcctggaag aggtggtgct ggttattcac
      481 gattggggca gcgccctggg attccactgg gccaagagaa accccgagag agtgaaggga
      541 atcgccttca tggaattcat caggcccatt cctacctggg acgagtggcc tgagttcgcc
      601 agagagacat tccaggcctt cagaaccacc gacgtgggca gaaagctgat catcgaccag
      661 aacgtgttca tcgagggcac cctgcctatg ggagtcgtca gaccactgac cgaggtggaa
      721 atggaccact acagagagcc ctttctgaac cccgtggacc gggaacctct ttggagattc
      781 cccaacgagc tgcccattgc cggcgagcct gctaatattg tggccctggt ggaagagtac
      841 atggactggc tgcatcagag ccccgtgcct aagctgctgt tttggggaac acccggcgtg
      901 ctgattcctc ctgctgaagc tgccagactg gccaagagcc tgcctaattg caaggccgtg
      961 gatatcggcc ctggcctgaa tctgctgcaa gaggacaacc ccgatctgat cggctccgag
     1021 attgccagat ggctgagcac cctggaaatc agcggataag tgttgatgta tcacctccgc
     1081 aaaactgttg gtaaatgtca gattttttcc tccaagagtt gtgcttttgt gttatttgtt
     1141 ttcactcaaa tattttgcct cattattctt gttttaaaag aaagaaaaca ggccgggcac
     1201 agtggctcat gcctgtaatc ccagcacttt gggaggtcga ggtgggtgga tcacttgggg
     1261 tcagggtttg agaccagcct ggccaacatg gcggaaccct gtctctacca aaattacaaa
     1321 aattagccga gcatggtggc gcatgcctgt agtcgcagct actcgcgagg ttgaggcagg
     1381 agaattgctt gaacccagga agtggcagtt gcagtgagcc gagacgacac cactgcactc
     1441 cagcctgggt gacagagggg gatcccgggc ccgtcgactg cagaggcctg catgcaagct
     1501 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1561 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1621 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1681 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     1741 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1801 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1861 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1921 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1981 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2041 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2101 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2161 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2221 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2281 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2341 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2401 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2461 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2521 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2581 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2641 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2701 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2761 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagatc
     2821 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2881 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2941 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3001 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3061 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3121 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3181 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3241 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3301 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3361 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3421 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3481 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3541 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3601 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3661 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     3721 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     3781 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     3841 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     3901 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     3961 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4021 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     4081 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     4141 tcggtacctc gcgaatgcat ctagatgcca ctgccctcca gcctgggtga cagcgagact
     4201 ccgtctcaaa aaaaaaaaaa aaaaaaaaaa atcctaaaat aatagggaag caggtatcac
     4261 ttggagagat ttttctctat gtgcatcgtg atgacttcag ttaaagacca aacacctgtg
     4321 ctcatgtccc actacgtgtt gaatacgaag ttgaactgat gttaaaactc gccatctgtt
     4381 cttcaagtga aacaaacaca actgcctgca aaatggaact aatggaatta tcatacttat
     4441 tc
//
