LOCUS       sequence-354942-        4473 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for endogenous tagging of human SLC1A3 at the 
            C-terminus with halotag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4473)
  TITLE     iNDI donor vectors for endogenous tagging with Halotag
REFERENCE   2  (bases 1 to 4473)
  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: SLC1A3_Halo_C_allele
FEATURES             Location/Qualifiers
     source          1..4473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             469..1356
                     /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(1818..1834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1818..1834)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1831..1853)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1842..1858
                     /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(1866..1872,1873..1890,1891..1896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1911..1932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2049..2066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2220..2808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2300..2319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2979..3770,3771..3839))
                     /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     3602..3621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3840..3944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4012..4030
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4068..4090)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4190..4209
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4403..4425
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4418..4434
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttccattcat catctatatg gccttataac ctttaatagc
       61 tactattatg aatattatgt gtctattacg taatcttgca gttggtagat acggcgaact
      121 gaatgttaag aggtgcttcg ctggccagtt cctaacgtaa gttgaaaact tgtgatgctc
      181 acgggagcct cgtttttccc tcctccccac cctgcctgca gggatcgcct ccggaccacc
      241 accaacgtac tgggagactc cctgggagct gggattgtgg agcacttgtc acgacatgaa
      301 ctgaagaaca gagatgttga aatgggtaac tcagtgattg aagagaatga aatgaagaaa
      361 ccatatcaac tgattgcaca ggacaatgaa actgagaaac ccatcgacag tgaaaccaag
      421 atgggtggag gaggttctgg aggcggtgga agtggtggcg gaggtagcgc cgagatcggc
      481 accggctttc ccttcgatcc tcactacgtg gaagtgctgg gcgagagaat gcactatgtg
      541 gacgtgggcc ccagagatgg aacccctgtg ctgtttctgc acggcaaccc taccagcagc
      601 tacgtgtggc ggaacatcat ccctcacgtg gcccctacac acagatgtat cgcccctgac
      661 ctgatcggca tgggcaagag cgataagccc gacctgggct acttcttcga cgaccacgtg
      721 cggttcatgg acgcctttat tgaggccctc ggcctggaag aggtggtgct ggttattcac
      781 gattggggca gcgccctggg attccactgg gccaagagaa accccgagag agtgaaggga
      841 atcgccttca tggaattcat caggcccatt cctacctggg acgagtggcc tgagttcgcc
      901 agagagacat tccaggcctt cagaaccacc gacgtgggca gaaagctgat catcgaccag
      961 aacgtgttca tcgagggcac cctgcctatg ggagtcgtca gaccactgac cgaggtggaa
     1021 atggaccact acagagagcc ctttctgaac cccgtggacc gggaacctct ttggagattc
     1081 cccaacgagc tgcccattgc cggcgagcct gctaatattg tggccctggt ggaagagtac
     1141 atggactggc tgcatcagag ccccgtgcct aagctgctgt tttggggaac acccggcgtg
     1201 ctgattcctc ctgctgaagc tgccagactg gccaagagcc tgcctaattg caaggccgtg
     1261 gatatcggcc ctggcctgaa tctgctgcaa gaggacaacc ccgatctgat cggctccgag
     1321 attgccagat ggctgagcac cctggaaatc agcggataaa ctaacataaa gaaacacttt
     1381 cttgagcacc aggtgttaaa aaccattata aaatctttcc atctcattac agctcattcg
     1441 ctccagcaag cccgtcatct tccctttcct cccttctgat aagactggaa aatagtcctc
     1501 caaaacacaa gggaggattt tgggtggcca aagtgtacaa ttttcatccc acaattgaaa
     1561 tttttaaatc atttcatgtt agtcttaccg aataaggtac caagatcaca aatagtgttg
     1621 atcagatctt acaagtttat gtggcacaca atcctataaa tgtgattttt ttatataagt
     1681 taaagagaca aatagtaggc taaaaacatt ttaaaatcaa cttttgaaat ttaaaaatct
     1741 ttcagaatac aattcagtta atcggatccc gggcccgtcg actgcagagg cctgcatgca
     1801 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1861 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1921 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1981 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     2041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     2101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     2161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     2401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     2461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     2521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     2581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     2641 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3121 gatccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3961 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     4021 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     4081 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     4141 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     4201 cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     4261 aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     4321 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     4381 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc
     4441 gagctcggta cctcgcgaat gcatctagat gtt
//
