LOCUS       sequence-368359-        5977 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Keratin labeling with blue fluorescent protein Electra2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5977)
  AUTHORS   Papadaki S, Wang X, Wang Y, Zhang H, Jia S, Liu S, Yang M, Zhang D, 
            Jia JM, Koster RW, Namikawa K, Piatkevich KD
  TITLE     Dual-expression system for blue fluorescent protein optimization.
  JOURNAL   Sci Rep. 2022 Jun 17;12(1):10190. doi: 10.1038/s41598-022-13214-0.
  PUBMED    35715437
REFERENCE   2  (bases 1 to 5977)
  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 Rep."; 
            date: "2022-06-17"; volume: "12(1):10190. doi"; pages: " 
            10.1038/s41598-022-13214-0"
COMMENT     Sequence Label: pKeratin-Electra2
FEATURES             Location/Qualifiers
     source          1..5977
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2790..2911
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2827..2846)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2881..2900
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2918..3373)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3055..3076)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3267..3286
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3400..3504
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3506..3863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3506..3697
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3529..3549)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3714..3849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3898..4692
                     /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             3898..4692
                     /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(3952..3971)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4562..4581
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4880..4899)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4924..4971
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5300..5888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5789..5808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact tttcaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag caccatgagc ttcaccactc gctccacctt ctccaccaac
      661 taccggtccc tgggctctgt ccaggcgccc agctacggcg cccggccggt cagcagcgcg
      721 gccagcgtct atgcaggcgc tgggggctct ggttcccgga tctccgtgtc ccgctccacc
      781 agcttcaggg gcggcatggg gtccgggggc ctggccaccg ggatagccgg gggtctggca
      841 ggaatgggag gcatccagaa cgagaaggag accatgcaaa gcctgaacga ccgcctggcc
      901 tcttacctgg acagagtgag gagcctggag accgagaacc ggaggctgga gagcaaaatc
      961 cgggagcact tggagaagaa gggaccccag gtcagagact ggagccatta cttcaagatc
     1021 atcgaggacc tgagggctca gatcttcgca aatactgtgg acaatgcccg catcgttctg
     1081 cagattgaca atgcccgtct tgctgctgat gactttagag tcaagtatga gacagagctg
     1141 gccatgcgcc agtctgtgga gaacgacatc catgggctcc gcaaggtcat tgatgacacc
     1201 aatatcacac gactgcagct ggagacagag atcgaggctc tcaaggagga gctgctcttc
     1261 atgaagaaga accacgaaga ggaagtaaaa ggcctacaag cccagattgc cagctctggg
     1321 ttgaccgtgg aggtagatgc ccccaaatct caggacctcg ccaagatcat ggcagacatc
     1381 cgggcccaat atgacgagct ggctcggaag aaccgagagg agctagacaa gtactggtct
     1441 cagcagattg aggagagcac cacagtgctc accacacagt ctgctgaggt tggagctgct
     1501 gagacgacgc tcacagagct gagacgtaca gtccagtcct tggagatcga cctggactcc
     1561 atgagaaatc tgaaggccag cttggagaac agcctgaggg aggtggaggc ccgctacgcc
     1621 ctacagatgg agcagctcaa cgggatcctg ctgcaccttg agtcagagct ggcacagacc
     1681 cgggcagagg gacagcgcca ggcccaggag tatgaggccc tgctgaacat caaggtcaag
     1741 ctggaggctg agatcgccac ctaccgccgc ctgctggaag atggcgagga ctttaatctt
     1801 ggtgatgcct tggacagcag caactccatg caaaccatcc aaaagaccac cacccgccgg
     1861 atagtggatg gcaaagtggt gtctgagacc aatgacacca aagttctgag gcatcgaatt
     1921 ctgcagtcga cggtacccat ggtgtctaag ggcgaagagc tgatcgagga aaatatgcgt
     1981 atgaaggtgg tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga
     2041 gaaggcagac cgtacgaggg agtgcaaacc atgaggatca aagtcatcga gggaggaccc
     2101 ctgccatttg cctttgacat tcttgccacg tcgttcctgt tcggcagcaa gacctttatc
     2161 aagtacccgg ccgacatccc tgatttcttc gaacagtcct ttcctgaggg ctttacttgg
     2221 gaaagagtta cgagatacga agatggtgga gtagtcaccg tcacgcagga caccagcctt
     2281 gaggatggtg ggctcgtcta caacgtcaag gtcagagggg taaactttca ctccaagggt
     2341 cccgtgatgc agaagaagac cgagggttgg gagcctttca cagagatgat gtatccagca
     2401 gatggtggtc tgagaggata taccgacatc gcactgaaag ttgatggtgg tggccatctg
     2461 cacgccaaca tcgtgacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc
     2521 ggtgtccatg ccgttgatta ccgcctggaa aggatcgaag agagtgacaa tgaaacctac
     2581 gtagtgctaa gagaagtggc agttgccaaa tacagcaacc ttggtggtgg catggacgag
     2641 ctgttcaagt aagatatcca gcacagtggc ggccgcgact ctagatcata atcagccata
     2701 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     2761 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     2821 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     2881 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     2941 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     3001 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     3061 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     3121 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     3181 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     3241 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     3301 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     3361 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3421 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3481 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3541 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3601 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3661 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3721 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3781 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3841 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     3901 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3961 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4021 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4081 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4141 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4201 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4261 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4321 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4381 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4441 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4501 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4561 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4621 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4681 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4741 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4801 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4861 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     4921 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     4981 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5041 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5101 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     5521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     5581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     5641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     5821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     5941 ctcacatgtt ctttcctgcg ttatcccctg attctgt
//
