LOCUS       sequence-491049-        5085 bp    DNA     circular SYN 29-OCT-2025
DEFINITION  anchoring mTFP1 to the chromatin/nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5085)
  AUTHORS   Vigano MA, Bieli D, Schaefer JV, Jakob RP, Matsuda S, Maier T, 
            Pluckthun A, Affolter M
  TITLE     DARPins recognizing mTFP1 as novel reagents for in vitro and in vivo
            protein manipulations.
  JOURNAL   Biol Open. 2018 Oct 29;7(11). pii: bio.036749. doi: 
            10.1242/bio.036749.
  PUBMED    30237292
REFERENCE   2  (bases 1 to 5085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 29, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biol 
            Open."; date: "2018-10-29"; volume: "7(11). pii: bio.036749. doi"; 
            pages: " 10.1242/bio.036749"
COMMENT     Sequence Label: pH2B-mTFP1
FEATURES             Location/Qualifiers
     source          1..5085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             657..1034
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     CDS             1041..1751
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    1873..1994
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1910..1929)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1964..1983
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2001..2456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2138..2159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2350..2369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2483..2587
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2589..2946
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2589..2780
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2612..2632)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2797..2932
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2981..3775
                     /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             2981..3775
                     /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(3035..3054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3645..3664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3963..3982)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4007..4054
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4383..4971
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4872..4891
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gccaccatgc
      661 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
      721 cgcagaagaa aggcggcaag aagcgcaagc gcagccgcaa ggagagctat tccatctatg
      781 tgtacaaggt tctgaagcag gtccaccctg acaccggcat ttcgtccaag gccatgggca
      841 tcatgaattc gtttgtgaac gacattttcg agcgcatcgc aggtgaggct tcccgcctgg
      901 cgcattacaa caagcgctcg accatcacct ccagggagat ccagacggcc gtgcgcctgc
      961 tgctgcctgg ggagttggcc aagcacgccg tgtccgaggg tactaaggcc atcaccaagt
     1021 acaccagcgc taaggatccg atggtgagca agggcgagga gaccacaatg ggcgtaatca
     1081 agcccgacat gaagatcaag ctgaagatgg agggcaacgt gaatggccac gccttcgtga
     1141 tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac ctggaggtga
     1201 aggagggagc ccccctgccc ttctcctacg acattctgac caccgcgttc gcctacggca
     1261 acagggcctt caccaagtac cccgacgaca tccccaacta cttcaagcag tccttccccg
     1321 agggctactc ttgggagcgc accatgacct tcgaggacaa gggcatcgtg aaggtgaagt
     1381 ccgacatctc catggaggag gactccttca tctacgagat acacctcaag ggcgagaact
     1441 tcccccccaa cggccccgtg atgcagaaga agaccaccgg ctgggacgcc tccaccgaga
     1501 ggatgtacgt gcgcgacggc gtgctgaagg gcgacgtcaa gcacaagctg ctgctggagg
     1561 gcggcggcca ccaccgcgtt gacttcaaga ccatctacag ggccaagaag gcggtgaagc
     1621 tgcccgacta tcactttgtg gaccaccgca tcgagatcct gaaccacgac aaggactaca
     1681 acaaggtgac cgtttacgag agcgccgtgg cccgcaactc caccgacggc atggacgagc
     1741 tgtacaagtg agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     1801 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     1861 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1921 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1981 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2041 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2101 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2161 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2221 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2281 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2341 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2401 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2461 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2521 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2581 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     2641 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     2881 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2941 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3481 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3541 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3721 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3781 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     3841 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     3901 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     3961 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4021 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4081 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4141 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4201 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4261 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4321 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4381 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4441 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4501 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4561 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4621 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4681 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4741 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     4801 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4861 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4921 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     4981 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5041 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
