LOCUS       sequence-160219-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses sfCherry2(11) tagged zyxin (human) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  AUTHORS   Feng S, Sekine S, Pessino V, Li H, Leonetti MD, Huang B
  TITLE     Improved split fluorescent proteins for endogenous protein labeling.
  JOURNAL   Nat Commun. 2017 Aug 29;8(1):370. doi: 10.1038/s41467-017-00494-8.
  PUBMED    28851864
REFERENCE   2  (bases 1 to 5757)
  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: "Nat 
            Commun."; date: "2017-08-29"; volume: "8(1):370. doi"; pages: " 
            10.1038/s41467-017-00494-8"
COMMENT     Sequence Label: pEGFP_sfCherry2(11)_zyxin
FEATURES             Location/Qualifiers
     source          1..5757
                     /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"
     regulatory      634..643
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2545..2666
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2582..2601)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2636..2655
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2673..3128)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2810..2831)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3022..3041
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3155..3259
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3261..3618
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3261..3452
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3284..3304)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3469..3604
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3653..4447
                     /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             3653..4447
                     /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(3707..3726)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4317..4336
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4635..4654)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4679..4726
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5055..5643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5544..5563
                     /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 attccatggc ggccccccgc ccgtctcccg
      661 cgatccccgt ttcggtctcg gctccggctt tttacgcccc gcagaagaag ttcggccctg
      721 tggtggcccc aaagcccaaa gtgaatccct tccggcccag ggacagcgag cctcccccgg
      781 cacccggggc ccagcgcgca cagatgggcc gggtgggcga tattcccccg ccgcccccgg
      841 aagactttcc cctgcctcca cctccccttg ctggggatgg cgacgatgca gagggtgctc
      901 tgggaggtgc cttcccgccg ccccctcccc cgatcgagga atcatttccc cctgcgcctc
      961 tggaggagga gatcttccct tccccgccgc ctcctccgga ggaggaggga gggcctgagg
     1021 cccccatacc gcccccacca cagcccaggg agaaggtgag cagtattgat ttggagatcg
     1081 actctctgtc ctcactgctg gatgacatga ccaagaatga tcctttcaaa gcccgggtgt
     1141 catctggata tgtgccccca ccagtggcca ctccattcag ttccaagtcc agtaccaagc
     1201 ctgcagccgg gggcacagca cccctgcctc cttggaagtc cccttccagc tcccagcctc
     1261 tgccccaggt tccggctccg gctcagagcc agacacagtt ccatgttcag ccccagcccc
     1321 agcccaagcc tcaggtccaa ctccatgtcc agtcccagac ccagcctgtg tctttggcta
     1381 acacccagcc ccgagggccc ccagcctcat ctccggctcc agcccctaag ttttctccag
     1441 tgactcctaa gtttactcct gtggcttcca agttcagtcc tggagcccca ggtggatctg
     1501 ggtcacaacc aaatcaaaaa ttggggcacc ccgaagctct ttctgctggc acaggctccc
     1561 ctcaacctcc cagcttcacc tatgcccagc agagggagaa gccccgagtg caggagaagc
     1621 agcaccccgt gcccccaccg gctcagaacc aaaaccaggt gcgctcccct ggggccccag
     1681 ggcccctgac tctgaaggag gtggaggagc tggagcagct gacccagcag ctaatgcagg
     1741 acatggagca tcctcagagg cagaatgtgg ctgtcaacga actctgcggc cgatgccatc
     1801 aacccctggc ccgggcgcag ccagccgtcc gcgctctagg gcagctgttc cacatcgcct
     1861 gcttcacctg ccaccagtgt gcgcagcagc tccagggcca gcagttctac agtctggagg
     1921 gggcgccgta ctgcgagggc tgttacactg acaccctgga gaagtgtaac acctgcgggg
     1981 agcccatcac tgaccgcatg ctgagggcca cgggcaaggc ctatcacccg cactgcttca
     2041 cctgtgtggt ctgcgcccgc cccctggagg gcacctcctt catcgtggac caggccaacc
     2101 ggccccactg tgtccccgac taccacaagc agtacgcccc gaggtgctcc gtctgctctg
     2161 agcccatcat gcctgagcct ggccgagatg agactgtgcg agtggtcgcc ctggacaaga
     2221 acttccacat gaagtgttac aagtgtgagg actgcgggaa gcccctgtcg attgaggcag
     2281 atgacaatgg ctgcttcccc ctggacggtc acgtgctctg tcggaagtgc cacactgcta
     2341 gagcccagac ggatccaccg gtcgccacca tgtacaccat cgtggagcag tacgagagag
     2401 ccgaggccag acacagcacc taagcggccg cgactctaga tcataatcag ccataccaca
     2461 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2521 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2581 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2641 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     2701 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     2761 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     2821 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     2881 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     2941 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3001 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3061 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3121 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3181 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3241 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3301 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3361 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3421 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3481 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3541 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3601 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3661 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3721 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3781 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     3841 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3901 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3961 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4021 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4081 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4141 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4201 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4261 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4321 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4381 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4441 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4501 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4561 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4621 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4681 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     4741 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4801 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4861 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     4921 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4981 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5041 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5101 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5161 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     5221 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5281 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5341 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5401 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5461 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5521 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5581 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5641 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5701 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
