LOCUS       sequence-243801-        5418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human SOX2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5418)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5418)
  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            "Unpublished"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-60: SOX2-mEGFP
FEATURES             Location/Qualifiers
     source          1..5418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..58
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(96..118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     218..237
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     431..453
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     445..462
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     446..462
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1516..1518,1519..2232)
                     /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(1558..1579)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1819..1838)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2166..2187
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3264..3280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3264..3280)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3277..3299)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3288..3304
                     /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(3312..3318,3319..3336,3337..3342))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3357..3378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3495..3512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3666..4254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3746..3765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4425..5216,5217..5285))
                     /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     5048..5067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5286..5390)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
       61 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      121 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      181 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      241 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      301 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      361 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      421 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgac gcgtattggg
      481 attgaattga cgcgtattgg gatgccgagg gtcggcggcc gccggcgggc cgggcccgcg
      541 cacagcgccc gcatgtacaa catgatggag acggagctga agccgccggg cccgcagcaa
      601 acttcggggg gcggcggcgg caactccacc gcggcggcgg ccggcggcaa ccagaaaaac
      661 agcccggacc gcgtcaagcg gcccatgaat gccttcatgg tgtggtcccg cgggcagcgg
      721 cgcaagatgg cccaggagaa ccccaagatg cacaactcgg agatcagcaa gcgcctgggc
      781 gccgagtgga aacttttgtc ggagacggag aagcggccgt tcatcgacga ggctaagcgg
      841 ctgcgagcgc tgcacatgaa ggagcacccg gattataaat accggccccg gcggaaaacc
      901 aagacgctca tgaagaagga taagtacacg ctgcccggcg ggctgctggc ccccggcggc
      961 aatagcatgg cgagcggggt cggggtgggc gccggcctgg gcgcgggcgt gaaccagcgc
     1021 atggacagtt acgcgcacat gaacggctgg agcaacggca gctacagcat gatgcaggac
     1081 cagctgggct acccgcagca cccgggcctc aatgcgcacg gcgcagcgca gatgcagccc
     1141 atgcaccgct acgacgtgag cgccctgcag tacaactcca tgaccagctc gcagacctac
     1201 atgaacggct cgcccaccta cagcatgtcc tactcgcagc agggcacccc tggcatggct
     1261 cttggctcca tgggttcggt ggtcaagtcc gaggccagct ccagcccccc tgtggttacc
     1321 tcttcctccc actccagggc gccctgccag gccggggacc tccgggacat gatcagcatg
     1381 tatctccccg gcgccgaggt gccggaaccc gccgccccca gcagacttca catgtcccag
     1441 cactaccaga gcggcccggt gcccggcacg gccattaacg gcacactgcc cctctcacac
     1501 atgggcggaa gcggagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1561 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1621 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1681 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1741 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1801 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1861 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1921 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1981 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2041 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2101 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     2161 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2221 ctgtacaagt gagggccgga cagcgaactg gaggggggag aaattttcaa agaaaaacga
     2281 gggaaatggg aggggtgcaa aagaggagag taagaaacag catggagaaa acccggtacg
     2341 ctcaaaaaga aaaaggaaaa aaaaaaatcc catcacccac agcaaatgac agctgcaaaa
     2401 gagaacacca atcccatcca cactcacgca aaaaccgcga tgccgacaag aaaactttta
     2461 tgagagagat cctggacttc tttttggggg actatttttg tacagagaaa acctggggag
     2521 ggtggggagg gcgggggaat ggaccttgta tagatctgga ggaaagaaag ctacgaaaaa
     2581 ctttttaaaa gttctagtgg tacggtagga gctttgcagg aagtttgcaa aagtctttac
     2641 caataatatt tagagctagt ctccaagcga cgaaaaaaat gttttaatat ttgcaagcaa
     2701 cttttgtaca gtatttatcg agataaacat ggcaatcaaa atgtccattg tttataagct
     2761 gagaatttgc caatattttt caaggagagg cttcttgctg aattttgatt ctgcagctga
     2821 aatttaggac agttgcaaac gtgaaaagaa gaaaattatt caaatttgga cattttaatt
     2881 gtttaaaaat tgtacaaaag gaaaaaatta gaataagtac tggcgaacca tctctgtggt
     2941 cttgtttaaa aagggcaaaa gttttagact gtactaaatt ttataactta ctgttaaaag
     3001 caaaaatggc catgcaggtt gacaccgttg gtaatttata atagcttttg ttcgatccca
     3061 actttccatt ttgttcagat aaaaaaaacc atgaaattac tgtgtttgaa atattttctt
     3121 atggtttgta atatttctgt aaatttattg tgatatttta aggttttccc ccctttattt
     3181 tccgtagttg tattttaaaa gattcggctc tgtattattt gaatcagtca tcccaatggc
     3241 gcgccgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3301 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3361 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3421 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3481 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5401 cgaaaagtgc cacctgac
//
