LOCUS       sequence-368347-        5994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Visualization of keratin using mGold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5994)
  AUTHORS   Lee J, Liu Z, Suzuki PH, Ahrens JF, Lai S, Lu X, Guan S, St-Pierre F
  TITLE     Versatile phenotype-activated cell sorting.
  JOURNAL   Sci Adv. 2020 Oct 23;6(43). pii: 6/43/eabb7438. doi: 
            10.1126/sciadv.abb7438. Print 2020 Oct.
  PUBMED    33097540
REFERENCE   2  (bases 1 to 5994)
  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: "Sci Adv."; 
            date: "2020-10-23"; volume: "6(43). pii: 6/43/eabb7438. doi"; pages:
            " 10.1126/sciadv.abb7438. Print 2020 Oct"
COMMENT     Sequence Label: pCMV-mGold-Keratin-N-17
FEATURES             Location/Qualifiers
     source          1..5994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..218
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     175..195
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(220..324)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      351..806
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(438..457)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     648..669
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    813..934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(824..843)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     878..897
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1056..1769,1770..1772,1773..1775))
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1101..1122)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1450..1469
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1709..1730
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1772..1781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3148..3351)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3177..3197)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3352..3655
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3830..4418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4747..4794
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4819..4838
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5026..5820)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(5026..5820)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5137..5156)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5747..5766
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5855..5994,1..218))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5869..5994,1..10)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc
       61 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat
      121 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta
      181 actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt ttcaatatta
      241 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
      301 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg
      361 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      421 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
      481 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
      541 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
      601 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
      661 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
      721 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
      781 taacaaaata ttaacgctta caatttacgc cttaagatac attgatgagt ttggacaaac
      841 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
      901 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
      961 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     1021 tggtatggct gattatgatc tagagtcgcg gccgcttact tgtacagctc gtccatgccg
     1081 agagtgatcc cggcggcggt cacgaactcc agcaggacca tgtgatcgcg cttctcgttg
     1141 gggtctttgc tcagcttgga ctggtagctc aggtagtggt tgtcgggcag cagcacgggg
     1201 ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga gctgcacgcc gccgtcctcg
     1261 atgttgtggc ggatcttgaa gttggccttg atgccgttct tctgcttgtc ggcggtgata
     1321 tagacgttgt ggctgttgta gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1381 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1441 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggacgtag
     1501 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcgggcg
     1561 aagcactgca ggccgtagcc caggctggtc acgagggtgg gccagggcac gggcagcttg
     1621 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcgccctcg
     1681 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1741 accccggtga acagctcctc gcccttgctc accatggtgg cgaccggtgg atcccgggcc
     1801 cgcggtaccg tcgactgcag aattcgatgc ctcagaactt tggtgtcatt ggtctcagac
     1861 accactttgc catccactat ccggcgggtg gtggtctttt ggatggtttg catggagttg
     1921 ctgctgtcca aggcatcacc aagattaaag tcctcgccat cttccagcag gcggcggtag
     1981 gtggcgatct cagcctccag cttgaccttg atgttcagca gggcctcata ctcctgggcc
     2041 tggcgctgtc cctctgcccg ggtctgtgcc agctctgact caaggtgcag caggatcccg
     2101 ttgagctgct ccatctgtag ggcgtagcgg gcctccacct ccctcaggct gttctccaag
     2161 ctggccttca gatttctcat ggagtccagg tcgatctcca aggactggac tgtacgtctc
     2221 agctctgtga gcgtcgtctc agcagctcca acctcagcag actgtgtggt gagcactgtg
     2281 gtgctctcct caatctgctg agaccagtac ttgtctagct cctctcggtt cttccgagcc
     2341 agctcgtcat attgggcccg gatgtctgcc atgatcttgg cgaggtcctg agatttgggg
     2401 gcatctacct ccacggtcaa cccagagctg gcaatctggg cttgtaggcc ttttacttcc
     2461 tcttcgtggt tcttcttcat gaagagcagc tcctccttga gagcctcgat ctctgtctcc
     2521 agctgcagtc gtgtgatatt ggtgtcatca atgaccttgc ggagcccatg gatgtcgttc
     2581 tccacagact ggcgcatggc cagctctgtc tcatacttga ctctaaagtc atcagcagca
     2641 agacgggcat tgtcaatctg cagaacgatg cgggcattgt ccacagtatt tgcgaagatc
     2701 tgagccctca ggtcctcgat gatcttgaag taatggctcc agtctctgac ctggggtccc
     2761 ttcttctcca agtgctcccg gattttgctc tccagcctcc ggttctcggt ctccaggctc
     2821 ctcactctgt ccaggtaaga ggccaggcgg tcgttcaggc tttgcatggt ctccttctcg
     2881 ttctggatgc ctcccattcc tgccagaccc ccggctatcc cggtggccag gcccccggac
     2941 cccatgccgc ccctgaagct ggtggagcgg gacacggaga tccgggaacc agagccccca
     3001 gcgcctgcat agacgctggc cgcgctgctg accggccggg cgccgtagct gggcgcctgg
     3061 acagagccca gggaccggta gttggtggag aaggtggagc gagtggtgaa gctcatggtg
     3121 ctagcggatc tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg
     3181 cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg
     3241 attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg
     3301 tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat
     3361 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     3421 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     3481 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     3541 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     3601 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     3661 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca
     3721 tggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4561 gtatatatga gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc
     4621 ctgggccttc acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt
     4681 ggccccaatg gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc
     4741 gtttatgaac aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag
     4801 cgcgggttcc ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg
     4861 cgaagaactc cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa
     4921 cgattccgaa gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca
     4981 ggttgggcgt cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc
     5041 aagaaggcga tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag
     5101 gaagcggtca gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat
     5161 gtcctgatag cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc
     5221 attttccacc atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc
     5281 gtcgggcatg ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc
     5341 ttcgtccaga tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat
     5401 gcgatgtttc gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg
     5461 cattgcatca gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc
     5521 ctgccccggc acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag
     5581 cacagctgcg caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg
     5641 cagttcattc agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc
     5701 tgacagccgg aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc
     5761 gaatagcctc tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat
     5821 gcgaaacgat cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa
     5881 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     5941 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gtta
//
