LOCUS       sequence-197677-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human SLC25A17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5406)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: AICSDP-27:SLC25A17-mEGFP
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1976..1994
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2032..2054)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2154..2173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2367..2389
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2381..2398
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2382..2398
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(3440..3442,3443..4156)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3482..3503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3743..3762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4090..4111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5188..5204)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5188..5204)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5201..5223)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5212..5228
                     /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(5236..5242,5243..5260,5261..5266))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5281..5302
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     1981 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     2041 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     2101 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     2161 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     2221 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     2281 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     2341 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     2401 attgacgcgt attgggataa atgtacaatt aaattattat tgactatagt cacctgttgt
     2461 gctagcaaat actaggtctt attcaaacta tctatttttg tacctattaa ccatccccac
     2521 cttccccccg ccactactct tcccagtagt ctctggtaac catccttcta cctttatctc
     2581 catgagttca attgttttga tttttaggtc cctcaaataa gtgagaacat gcgatatttg
     2641 tctttctgtg cctggtttgt ttcacttagc agaatgacct ccagttccat ccatgttgtt
     2701 acaaacaaca aactctcatt ctttttgatg gctgaatagt actgcatttt gtataagtac
     2761 cacattttct ttatccattt atctgttgat ggacatgtag cttgcttcca aatttaagac
     2821 attatttgta agaaccaaaa actagggcct ggcacggtgg ctcacacctg taatcccagc
     2881 actttgggag gctgaggcgg gcagatcacg aggtcagggg atcgagacca tcctggctaa
     2941 catggtgaaa ccccatctcc actaaaaata caaaaaaaaa ttagcctggt gtcgtggcgg
     3001 gtgcctctag tcccagctgc tagggaggct gaggcaggag aatggcatga actcgggagg
     3061 cggagcttgc agtgagccga gatcgcgcca ctgcactcca gcctgggcga cagagcgaga
     3121 ctccgtctca aaaaaaggaa agtactttga caacagaagt ctgtgttgaa atctaaaacc
     3181 tctattgctt gttcttatca aagtagctat ctaggtagca tgttctctga tgcaggagct
     3241 gacttctgtt ttttcaaacc tctttccctt tagacgtttt ggaataatgg gactctacaa
     3301 aggccttgaa gccaaactgc tgcagacagt cctcactgct gctctcatgt tccttgttta
     3361 tgagaaactg acagctgcca ccttcacagt tatggggctg aagcgtgcac accaacacag
     3421 agacccccct gtcgccaccg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     3481 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3541 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3601 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3661 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3721 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3781 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3841 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3901 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3961 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4021 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     4081 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4141 cgagctgtac aagtgagacg gcttgccatg aaaaattccg aagatgctca agagggaggt
     4201 ttcctcctga gtgaagagaa gtgattctcc cttgactctg gctcctgcca ccacaaatgt
     4261 taccctcatt ggcttgaaaa gcatccaagg gtgcacaggg agtatggcca actggacctg
     4321 ttgtcacctt aattgtcatg ctggctggtt ggattttggg gtggcagttg gactaatgtg
     4381 aaaaaaacat tgctgaaaac ctaaaaatga aagtttgtga gtgtttattg gttttcttaa
     4441 gagaaatgga ctattttgct ctcatgtgta atgttttcta tttaaatctt tcttaaatat
     4501 accagctgtt ctctttccct gaactctccc ccaggttcta ggacaaattt aataacatgt
     4561 aattctcctc aaatactttt gtatgtctca gtgttggtgt tttcctccct aaaactaaca
     4621 ttagggctgt gccacgggca tgactttatt tttgttgggc ttttttttcc ctgcttaagg
     4681 agaggtgtct tttttggata tgagctattt attttgtgaa atgaaaattg ttcacccaaa
     4741 tgattctctt ataaactatt tgtaaatgtc acttattcat tagtgtttga cataattttt
     4801 agaatattta ttttgaatca atcctttcat tacgaaagac ttgaagtttt gtgtccattc
     4861 ttacaagccc tggtcagtca agtcccaata aatggtcagc acaaaaaaga tattcttgaa
     4921 aattgctctt tattaaggta ttacgttgag tttgcaacca gatgggaaaa atcacaaaaa
     4981 tgagaagggg agcagatatc ttgttgaggt ctggatatta tctcccttta taaacttggt
     5041 gtaggccgta tattttgaaa ataaacatct ggttgaggtt atttcatttg gaaaccttcc
     5101 ttagagtcct gttgctcagt gtggtccatc agacggtagt actgccacaa gctatcccaa
     5161 tggcgcgccg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5221 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5281 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5341 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5401 tgggcg
//
