LOCUS       sequence-220871-        5397 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mEGFP-linker sequence for N-terminus tagging of 
            human CBX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5397)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5397)
  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            "Unpublished"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-59: CBX1-mEGFP
FEATURES             Location/Qualifiers
     source          1..5397
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..72)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     172..191
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     385..407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     399..416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     400..416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1434..1436,1437..1439,1440..1442,1443..2153)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MLVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD
                     DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGI
                     KVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVL
                     LEFVTAAGITLGMDELYK"
     primer_bind     complement(1482..1503)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1743..1762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2090..2111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3197..3213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3197..3213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3210..3232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3221..3237
                     /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(3245..3251,3252..3269,3270..3275))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3290..3311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3428..3445)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3599..4187)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3679..3698)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4358..5149,5150..5218))
                     /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     4981..5000
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5219..5323)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(5391..5397,1..12)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
       61 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
      121 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
      181 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
      241 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
      301 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
      361 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
      421 tgacgcgtat tgggatttgt atttttagta gggacagggt ctcgccatgt tggccaggat
      481 ggtcttgaac ccctgacctc aggtgatctg cccgcctcgg cctcccaaag tgctagaatt
      541 ataggtgtga gccactgcac ccagcctttc aatttaatgc aatattttag ctaccgtgct
      601 cttttacaac ggtttttttg tgaagatgga gtcttgctgt gttgcctagg ctggtcacac
      661 actactgggc ttcaggaatt ctcccacttg ggccttccaa aatgctggga ttgcagatgt
      721 gagcctgcac gcctggccct agctaccatg ctttatggta aaaggagtat tgggagcctt
      781 ttaaaattac caagaatggg tataaaagtt ttaaatggat acacaagaat aaagaagttt
      841 gctgtgaagg ccaggcgtgg tggctcatgc ctgtaattcc aacactttgg gaggccgagg
      901 caggtggatc acctgaggtc aggagttcga gaccaacctg gccaacacat agtgaaatcc
      961 cgtctctact aaaaaaaaaa aaaaaacaaa aacaaaaaaa aaaacaacaa aaattagctg
     1021 ggcttggtgg cacacgcctg tagtcccagc tgcttgggaa gctgaggcag gagaatcgct
     1081 tgaacccagg aggcggaggt tgcagtaagc cgagattgcg ccactttgca ctccagcctg
     1141 ggcaacagag caagactccg tctctcaaaa aaaaaaaaaa agtttgctgt aagattaatt
     1201 ggggctgttt tatgagcttt ctggctggac atatcagaaa actattttgt atcgagtgtg
     1261 agaagaagcc taacacttac tataagcagc atacaatatt atcacctgtc atgtccctgg
     1321 ttttcttacc atccagatat tatctaacca ataagagtgc tccctaatgc cctttttatt
     1381 tcatttatca ttttagcagc gtcagccttt acaccagaaa gctggcgggc actatgctgg
     1441 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1501 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1561 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1621 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1681 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1741 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1801 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1861 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1921 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1981 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     2041 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     2101 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagaagggca
     2161 gcgggaaaaa acaaaacaag aagaaagtgg aggaggtgct agaagaggag gaagaggaat
     2221 atgtggtgga aaaagttctc gaccgtcgag tggtaaaggg caaagtggag tacctcctaa
     2281 agtggaaggg attctcagag taagtttcac tgacacaggt aaatgtagcc accacgtgtt
     2341 tatcaggagt ctagagatgt atgagacctc cagtgctatg atggatacag tgtgactcag
     2401 tccccacctt catggcttat tgtttaatta ggaatatggc ctactgctta taatacaaag
     2461 cagaatgact tctttttaaa aattatttct tttgagacga ggtctcgctc tgtcacccag
     2521 gctggagtac agtggtgcag tcatagctcg ctgcagcctc aacctcccag gctcaaggta
     2581 tcctcccacc tcagccttct aagtagctgg taccacaggc atgcaccacc atgtctcgct
     2641 tagttgccca ggtttgtctc ttaactcctg gcctcaagca gtcctcccac ctgggcctcc
     2701 caaagtgttg ggactacaag catgagccac tgtacccagc ccatagtgat tgctgagaaa
     2761 cagttttgtg tgtggctata gaccaattta aattgacaaa taaaagtcat ttgcctttga
     2821 aggaagccag tttcttagct ttctatttat actaaaaagc ttccccattc cccaagaatt
     2881 tcagcttttc catagacatc tgaggtcttg tcctcttacg attatcctta gtatgagtat
     2941 acttatttga gtgaaagtgt gacttgctgt gccctctggt ttcagtgagg acaacacatg
     3001 ggagccagaa gagaacctgg attgccccga cctcattgct gagtttctgc agtcacagaa
     3061 aacagcacat gagacagata aatcagaggg aggcaagcgc aaagctgatt ctgattctga
     3121 agataaggga gaggagagca aaccaaagaa gaagaaagaa gaatcccaat ggcgcgccga
     3181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3781 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4021 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5341 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggc
//
