LOCUS       sequence-220830-        5394 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human ATP2A2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5394)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5394)
  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            "Unpublished"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-41: ATP2A2-mEGFP
FEATURES             Location/Qualifiers
     source          1..5394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     227..249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     241..258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     242..258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1276..1278,1279..1281,1282..1992)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1321..1342)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1582..1601)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1929..1950
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3036..3052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3036..3052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3049..3071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3060..3076
                     /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(3084..3090,3091..3108,3109..3114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3129..3150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3267..3284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3438..4026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3518..3537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4197..4988,4989..5057))
                     /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     4820..4839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5058..5162)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5230..5248
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5286..5308)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
       61 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
      121 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
      181 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
      241 tgtaaaacga cggccagtga attgacgcgt attgggatcg cagcccgggc gaccgagggc
      301 gaggaggcga gccaaggaca tcagcccgag ggcgcctcga gacgccccgc gtggaccgcg
      361 ctcccagctc ctcggcctcg ccttccaacc atccgcccac cggccccaga gcagcgtgcc
      421 cactgtgagc gccccaccct gcgtctgcag gtgggtgggt cagagaaccg caggcacaga
      481 agagggtacc cagcttcccc tccgccagcc ccgcgaccgc ggcgcgcgcg gcctcgatcc
      541 gggttcctag gggcggcgcg cgggaggggg cggggcctgc gcggcagcgt gggcgccagg
      601 cgcgcgggag gagggagccg ggaggagggg gcggggccgc gccgcccgcg ccgcgctggg
      661 cgctctcggc caatgagcgg cgtccacatg ccgcggcggc ggcgaaaggg gaggcagcgg
      721 ccgataaatg ctattagagc agccgccgcg gagccgtccc cgacgccacc tccttttcct
      781 tcgccgcagt ttcctccgcc gctgtcgggc gtgcggcgct gagggacccg ggcgagcgcg
      841 ccgcgcaccg ccccgccggc tcgcctccct cgccgcgttc cgccctcagt ggtctgccgg
      901 gcgccccctc ctccggcccg ggcggggcct ctgatcgcct caagagagcg gggagggggc
      961 tcgggggccg cggcctgccc tcccggcggg cggctgaggg cgagggaggc cctcccttct
     1021 ggcgagggga gggagggtgg gtcaggagcc cccaacccgc cctgcggagc tcggggccgc
     1081 gcgaggggcg gttgtctggg ggagggggcg cggggtgatt cagcgcccgg cgaggcggaa
     1141 gcggccgcaa gaggaggagg ggagagcccg tccgcgcctg ggctcccggg gtggcacgag
     1201 cccgcggccg gagtgcgagg cggaggcgag gaggccgcgg ggacgggagg cgaggccggc
     1261 cgggcccccg aagccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1321 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1381 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1441 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1501 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1561 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1621 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1681 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1741 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1801 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1861 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     1921 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1981 gagctgtaca agggcagcgc tgagaacgcg cacaccaaga cggtggagga ggtgctggga
     2041 cacttcggcg tcaacgagag tacggggctg agcctggaac aggtcaagaa gcttaaggag
     2101 agatggggct ccaacggtag gtgcagggcg ctccgctgca ggggcccggc gcggccggga
     2161 gagccaggga agatggctga ccgggctcca cctcgtgggc ttcggctccg cgcccgccga
     2221 cagctgcggg cggagggtcg ggccagcgcg ccggccccgc gggagagaaa ggggctgcgg
     2281 tcctcgcctc gccttccctg gacctcttcg cttctcgggc cctcgacctt ttcggcgcgc
     2341 aaggctcgga ggcttctctc cagcagcagc cggccccggt ggagggaggg acgtggcttc
     2401 tgcagctagg ttgagcccgg caagacgttt tctcgtcccc tgccgattta tgaggagtcg
     2461 aggtcgttgg aaggcctctg accgttcttg tcctacccaa agttacacat ctggcagaag
     2521 tgatgacatc gctgaaacca ctcctaggtt caggagcccg aagtgatttc acgcttaggg
     2581 ctagacctca ggccattgat tacaaagagt gcatgcttcc ttgagatatc ttcgtggtat
     2641 gccctctgct aaaaaaatcg gatattctta tagctaggtc tgtgttttaa agatattgat
     2701 gcttagaatt gtagcagttc tttttagaaa aacgaagtgc taaaatgtct ctctcttttt
     2761 ttttttaacc tccctcttga cacattgctt gacgaatttc tacattctac agagttaccg
     2821 gctgaagaag gtaatcttaa catgctgttt ctgttttttt tcctctgttg gtgtgctgat
     2881 ggtaagatga cagttaaaac acatgtgttt gtttcttaca ggaaaaacct tgctggaact
     2941 tgtgattgag cagtttgaag acttgctagt taggatttta ttactggcag catgtatatc
     3001 tatcccaatg gcgcgccgag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagtggcggt
     3241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     3781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     3841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     3901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     3961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4141 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4201 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4261 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4321 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4381 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4441 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4501 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4561 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4621 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4681 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4741 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     4801 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     4861 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     4921 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     4981 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5041 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5161 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5221 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5281 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5341 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggc
//
