LOCUS       sequence-220833-        5412 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for C-terminus tagging of 
            human SMC1A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5412)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5412)
  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            "Unpublished"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-57: SMC1A-mEGFP
FEATURES             Location/Qualifiers
     source          1..5412
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     23..40
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     24..40
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1088..1090,1091..1804)
                     /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(1130..1151)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1391..1410)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1738..1759
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2836..2852)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2836..2852)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2849..2871)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2860..2876
                     /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(2884..2890,2891..2908,2909..2914))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2929..2950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3067..3084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3238..3826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3997..4788,4789..4857))
                     /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     4620..4639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4858..4962)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5030..5048
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5086..5108)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5208..5227
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgacgc gtattgggat
       61 ctactggttc cctcccagct acaagccagc ccccttcttc gtcctggatg agattgatgc
      121 tgccttggat aacaccaaca ttggcaaggt gggtgcaaag aaagtggggc ctgggaggga
      181 tgcccccagg tcggggttgg gtttcaggga gcaactcctg agcgtgccct agccgttcct
      241 gctgcctgct ctctgtgcct gggaggctgg gcccagagtc atccatgctg tggtagtcag
      301 tagtctcctc tcaacctctc ttttccccac cttcctctcc ctaccctacc tcttctgcct
      361 tctggttgtg gctggaaaat actggggact gttaaggttt gagccttgct aagtgcctgg
      421 cggcctttcc cacaggtggc aaattacatc aaggagcagt cgacttgcaa cttccaggcc
      481 atcgtcatct ctctcaagga ggagttctac accaaggccg agagcctcat tggagtctat
      541 cctgaggtag ggcgggcctg gctcaggagc cagtcctact ccctgcctga tttcccaggg
      601 caggaaagga aggctgcagt ggaggggaaa ggaggtgggg gagcataggg aatcaggtcc
      661 tgaatggcca tttgggcttg ttggagccct gcctctggag gcttagccag cccagccagg
      721 cagggccatt gcatttgggg acaggtggaa aggcaggcag aggcagccag acagtggaga
      781 aaggaggaga gtacagaagt aggcaaggag cgaaggcaca tgggactgag catatcctct
      841 cacacccaga gagaaccatt gactgggctt tgggcaggta tgtagggagg agggtttgag
      901 gcccctggtc ctggggcact agacccctct taattctctc cttacaattt tatttttctt
      961 tttctccctc ctccttcagc aaggggactg tgtgatcagc aaagtcctga ccttcgacct
     1021 caccaagtac ccagatgcca accccaaccc caatgagcag ctgagctcca gcggcccctc
     1081 aggcagcgtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1141 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1201 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1261 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1321 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1381 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1441 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1501 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1561 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1621 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1681 ccactacctg agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat
     1741 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1801 gtagcagtat ttttgtcctc cggccctgtc tggatcccta agctgtccct ctcccaatct
     1861 ctggatattt gactcccaac cttcccccta cctcctggcc ctttttggtg tagtcatggg
     1921 atttaggcac tgctaatcaa gcatgaagag gaacagaggt gatgttaggt ctggagcaaa
     1981 aattcctgaa cgacagggag tattctggcc tctgaaagga ggtgctgagc tgaacagggc
     2041 catctgttca tcacacacac ccccttcctc cccctcatca cccataatcg tgggcccctt
     2101 gggcctcttg cccactgtgt gtgtgggtat gtatgtgtgt atgtatgtat ccgcatgtgt
     2161 gcatgtgagt atgtttgcaa aataataaag gatattggag acctgtttta gaaggagcct
     2221 aggctgaatt tgattccaag agagcttagg atgacagcac ccctgagctg ggcaaaggta
     2281 ctcaggacct cataggagtc ttaggcagtt acctgaaact gccttcattc actcatttgt
     2341 gtattcattc atttatgtat tcatcagaca cataccgaac accctctatt tgtcaggctc
     2401 tgtgcttgga atacagagtt gaatcagaca tgatctctac cctcctagta aggagataca
     2461 gtgggttcat gaatgactat agttagctga atgtcatatg tactttgaat ttgagaagtg
     2521 ggtgatcccc tctaggcttc ctggaggtca catttaagct agaccttgac aaattggtag
     2581 gatttggtca ggcactagga gtggagcatg agctctgggg acagacagtt atgggttctg
     2641 gtcccacttt ttatcactta ctagttgttt gaccttgggc aagtcatttg accttctgtg
     2701 cctcagtttc ctcatctgta aaatggggct aacaatatta cctacctcat aggatttaat
     2761 gatgtcaagc tcctcactgg aggccttatc ccttcgtgga gatcccaatg gcgcgccgag
     2821 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2881 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2941 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3001 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4981 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5041 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5101 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5161 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5221 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5281 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5341 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     5401 ttgggtaacg cc
//
