LOCUS       sequence-173967-        5351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and linker-mEGFP sequence for N-terminus tagging of 
            human SEC61B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5351)
  AUTHORS   Roberts B, Haupt A, Tucker A, Grancharova T, Arakaki J, Fuqua MA, 
            Nelson A, Hookway C, Ludmann SA, Mueller IA, Yang R, Horwitz R, 
            Rafelski SM, Gunawardane RN
  TITLE     Systematic gene tagging using CRISPR/Cas9 in human stem cells to 
            illuminate cell organization.
  JOURNAL   Mol Biol Cell. 2017 Oct 15;28(21):2854-2874. doi: 
            10.1091/mbc.E17-03-0209. Epub 2017 Aug 16.
  PUBMED    28814507
REFERENCE   2  (bases 1 to 5351)
  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"; doi: 
            "10.1091/mbc.E17-03-0209"; journalName: "Mol Biol Cell"; date: 
            "2017-10-15- 15"; volume: "28"; issue: "21"; pages: "2854-2874"
COMMENT     Sequence Label: AICSDP-7:SEC61B-mEGFP
FEATURES             Location/Qualifiers
     source          1..5351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(1413..1415,1416..1418,1419..2129)
                     /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(1458..1479)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1719..1738)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2066..2087
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3130..3146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3130..3146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3143..3165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3154..3170
                     /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(3178..3184,3185..3202,3203..3208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3223..3244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3361..3378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3532..4120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3612..3631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4291..5082,5083..5151))
                     /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     4914..4933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5152..5256)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5324..5342
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggattttaa
      421 atgggcccac actaaagtta gagaaccaca ggctcgctca caaccctgac ttctccatgt
      481 cagttccgat ctttgcgaac cgcagacagg gaaggtcttc tctcaggggt catgcccgcg
      541 gccgccctcc acggcgaggt ccgcactcgc gcagccggcc ccgcggccgc ctcacctggt
      601 cgcacactac cacgtcgaac tcctcgtcgg cgaggaacag cacgtagagc gccaggaaaa
      661 ccatgcgcac gtaggcgcag acggcggcgc cgcggccgcc ccagcccagg cctcgcggca
      721 gccagtcccc ggcacagcgc accggtagct cgcggctctc ggcgaaacag tggcccgggt
      781 cgtagtgcgc tgtccagatc ttcacgctac acccgcgcgc ctgcagcgcc agcgccgcgt
      841 ccaacaccag ccgctcagcg ccgcccacgc ccaggtctgg gtggaggaac agcaccgacg
      901 gcttgggaac cgagtcccgt tcccggccct gctcctccgc catggccctg gagccgcaac
      961 tgcaccccgc accctgatgg gggtcttctg cgcaagctcc gcgctcgtag ctcccagctg
     1021 gccactgcgg gccgaccccg ccctgccgta cgtgcgtcag ttaggccaca tcagcgcaaa
     1081 tctgtgaggg tctagtaact gcctgagaaa atatcttgtc tgaccccggt tatatttttc
     1141 cttcggtagg gattggactt tctgaaggac gttgtgatcc aaaggaagga ggccggaggt
     1201 ctctacttcc catacagcag gtaactaagt tgtctgtagc agactgtcta caggcatatc
     1261 gtgagacgac ccaggcgtcc ctggggtcag agaggacctt gcctgcaagt ccgggggcgg
     1321 ggcctgagtc agtctcgcca gctgccggtc tttcgggggc tccgtaactt tctatccgtc
     1381 cgcgtcagcg ccttgccaca ctcatctcca atatggtgag caagggcgag gagctgttca
     1441 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1501 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1561 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1621 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1681 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1741 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1801 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1861 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1921 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1981 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     2041 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     2101 tcactctcgg catggacgag ctgtacaaga gcggcctgag aagcatggta tggcggccct
     2161 tccatgatcc ccgcctctcc cagaagccct gactcctcct gctttgcgcc gtgcttttcc
     2221 tctgtagctc ccttgcttcc cccagcctcg ggtgtgggtg tctaggccgg ggttctgggg
     2281 caggcctgcc gcgctcaccc gtctgtctgc ttgtctccct ctacagcctg gtccgacccc
     2341 cagtggcact aacgtgggat cctcagggcg ctctcccagc aaagcagtgg ccgcccgggc
     2401 ggcgggatcc actgtccggc agaggtaagg aaccctgcag ttcgttcgct tccagactcg
     2461 gagataggac ccagaacctc gctgattctg gggtggagac cctagcatgt gaagattgac
     2521 aaaggcaaaa tgagcttcta gtgacgtggc cgtgggagta gttaaaggcc ttttgggagg
     2581 aaggcgacat tttttttctc gttgctcagt ttagggcact actcttaaaa aaggaaagtt
     2641 aacaaactgg aatagagtca gagataactt tgagaaaacc gatgtcatta aactggtgtc
     2701 tctggacctg aggtttgcac tcacatttcc atctggcggc cccataagca atctgtccta
     2761 cagataactc gtcctacaca aaacttagtc tcttttcagc tcagctctct cactctcaat
     2821 tatatctcct tacttccata tggcactgtt gtacactcat ttactcagag ccagaaacgt
     2881 cagcgtcatc ttggattttt cttatgctct ttctctctct agtcatatgc cagactttaa
     2941 actctgcttg aaagctttct cataagctct ttccttttcc ctttctactg ctttgcattt
     3001 gctacttaac ccttttcttc aggctgtttg ctttccagtc catcgttcgc tctgctgtta
     3061 ctcttctgcg tagtttctgt tacttgttgc tgaacatccc aatggcgcgc cgagcttggc
     3121 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3181 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3241 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3301 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3361 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3421 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3481 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5161 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5221 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5281 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5341 gccctttcgt c
//
