LOCUS       sequence-349345-        4374 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for endogenous tagging of human SGP11 at the N-terminus
            with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4374)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: SGP11_Halo_N_allele
FEATURES             Location/Qualifiers
     source          1..4374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(576..592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(576..592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(589..611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    600..616
                     /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(624..630,631..648,649..654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    669..690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(807..824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(978..1566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1058..1077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1737..2528,2529..2597))
                     /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     2360..2379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2598..2702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2770..2788
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2826..2848)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2948..2967
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3161..3183
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3175..3192
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3176..3192
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3599..4374,1..115)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
ORIGIN
        1 gcctaattgc aaggccgtgg atatcggccc tggcctgaat ctgctgcaag aggacaaccc
       61 cgatctgatc ggctccgaga ttgccagatg gctgagcacc ctggaaatca gcggaggagg
      121 aggttctgga ggcggtggaa gtggtggcgg aggtagcgct gcagaggaag gggtcgcgag
      181 tgctgcttcc gccggcggta gctggggcac cgcggccatg gggcgggttc taccgatgct
      241 gttggtgcca gtccccgccg aggcgatggg gcagctcggc tcccgggcgc agctgcgcac
      301 acagccggag gctctgggga gcctgacggc tgcgggcagc ctccaagtgc tttctttgac
      361 gcctggcagc cggggcgggg gtcgctgctg cctggagggc cccttctggc agtaagtgct
      421 gagggagagt tgggccgttg ggggagtcca gagcctaggc tcggctgact gagccccaag
      481 ggctcacgcc gaggcctccc tgcagccata gaggtggcgg gagaagggtg ccagctgcag
      541 gaaacgcccc agaggcgaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      601 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      661 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      721 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      781 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      841 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      901 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      961 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     1021 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1081 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     1141 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     1201 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     1261 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     1321 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     1381 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     1441 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1501 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1561 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1621 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1681 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1741 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1801 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1861 gctgcaatga taccgcgaga tccacgctca ccggctccag atttatcagc aataaaccag
     1921 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1981 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     2041 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     2101 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     2161 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     2221 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     2281 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     2341 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2401 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2461 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2521 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2581 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2641 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2701 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2761 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2821 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2881 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2941 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     3001 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     3061 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     3121 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     3181 acgacggcca gtgaattcaa ggaaatggca ccaccccaga gcctgtaagc gaacagcaca
     3241 ataagaaaag cgaagaaaag aggcaggaac tgggttccaa gggagctgga ggcagctgga
     3301 ccaagaaggc aaagagcaat gccacagagc gagaacggct gcaacggcct gctacgctaa
     3361 gctaggcctt caagcatgcc agagcagtta agcagagtgg gactggcgtc agggcagatg
     3421 tggctttgct agagaggctg gtactgcccc ccacaggaaa cgaatggaat cgaccggaga
     3481 cagccgagag gtctccgcgg gagtcacgag gccgagcggg agccgctacg tacaccgcgc
     3541 atgcccacga cgcagtcagg ttccggcgaa agtgaccgga agtaaccgcc gggccaagat
     3601 ggccgagatc ggcaccggct ttcccttcga tcctcactac gtggaagtgc tgggcgagag
     3661 aatgcactat gtggacgtgg gccccagaga tggaacccct gtgctgtttc tgcacggcaa
     3721 ccctaccagc agctacgtgt ggcggaacat catccctcac gtggccccta cacacagatg
     3781 tatcgcccct gacctgatcg gcatgggcaa gagcgataag cccgacctgg gctacttctt
     3841 cgacgaccac gtgcggttca tggacgcctt tattgaggcc ctcggcctgg aagaggtggt
     3901 gctggttatt cacgattggg gcagcgccct gggattccac tgggccaaga gaaaccccga
     3961 gagagtgaag ggaatcgcct tcatggaatt catcaggccc attcctacct gggacgagtg
     4021 gcctgagttc gccagagaga cattccaggc cttcagaacc accgacgtgg gcagaaagct
     4081 gatcatcgac cagaacgtgt tcatcgaggg caccctgcct atgggagtcg tcagaccact
     4141 gaccgaggtg gaaatggacc actacagaga gccctttctg aaccccgtgg accgggaacc
     4201 tctttggaga ttccccaacg agctgcccat tgccggcgag cctgctaata ttgtggccct
     4261 ggtggaagag tacatggact ggctgcatca gagccccgtg cctaagctgc tgttttgggg
     4321 aacacccggc gtgctgattc ctcctgctga agctgccaga ctggccaaga gcct
//
