LOCUS       sequence-433847-        6131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express mCherry-AHPH (RhoA biosensor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6131)
  AUTHORS   Budnar S, Husain KB, Gomez GA, Naghibosadat M, Varma A, Verma S, 
            Hamilton NA, Morris RG, Yap AS
  TITLE     Anillin Promotes Cell Contractility by Cyclic Resetting of RhoA 
            Residence Kinetics
  JOURNAL   Dev Cell. 2019 Jun 17;49(6):894-906.e12. doi: 
            10.1016/j.devcel.2019.04.031. Epub 2019 May 16.
  PUBMED    31105010
REFERENCE   2  (bases 1 to 6131)
  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: 
            "10.1016/j.devcel.2019.04.031"; journalName: "Dev Cell"; date: 
            "2019-06-17- 17"; volume: "49"; issue: "6"; pages: "894-906.e12"
COMMENT     Sequence Label: pmCherry-AHPH
FEATURES             Location/Qualifiers
     source          1..6131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..330
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     287..307
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(332..436)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      463..918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(550..569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     760..781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    925..1046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(936..955)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     990..1009
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2920..2939)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3189..3207
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3349..3358
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3397..3600)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3426..3446)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3601..3904
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4079..4667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4159..4178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4996..5043
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5068..5087
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5275..6069)
                     /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(5275..6069)
                     /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(5386..5405)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5996..6015
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6104..6131,1..330))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6118..6131,1..122)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cctcctcact acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata
       61 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
      121 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
      181 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
      241 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
      301 acattccaca gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca
      361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
      481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      901 tattaacgct tacaatttac gccttaagat acattgatga gtttggacaa accacaacta
      961 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     1021 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     1081 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg
     1141 ctgattatga tcagttatct agatccggtg gatcccgggc ccgcggtcaa ggctttccaa
     1201 taggtttgta gcaagcatca ggttgccaga ggcgaatatc aacaagaact tgattgagtt
     1261 tttgcatcca gagatcccgc tcttctttag tatctgcaga cagccagttc ttggtaacac
     1321 agagtgtgtc cctgcattgg ctgacaagag tctctcggtc atcttctctt tgtggtcgga
     1381 cagtaattaa ttcaaaagtg ttgcgtcttg cacaaaattc tctgttggct ggttctatct
     1441 gacgactggt acaattagcc agatttatcc ttcctatggg attcttgcgt ttctcatcat
     1501 ctggataagt ccaataagat atacagtttc cagaaagaac acaccatctt cgatgccagg
     1561 caccaaaacc actaacatct tcaaatatgg ttagaaaacc tctttcttca acactggaat
     1621 tcacttgaca ttttattttt aaataaatat gaccttccaa agaagataaa aaggggacct
     1681 tgtccagaac aaacttagta tttcctactg aagacaatga taatgtgtaa gatccaacaa
     1741 gggcgaagtt gctggttcgc acagcactaa gacctcctgg actggccatg actgaagaat
     1801 gaatgttgct ttttgtggtt atagatgtga ggagtcgctt tggagtaata gccttggact
     1861 tggatgtttt tttcttctta tcaaggcctg agggatcttt cttttgcacc aagctgtaaa
     1921 cttcaatatt tatttcaaag tcattggata catcttgcag agtaaatgta gtagtgaatg
     1981 tcagagcatc accgttaaga gagtttgaag tacttgctaa tggtgtggct accatatttt
     2041 cagctcctgc ttttagtata attaagtaat agtaatttgc tgcatctggt ttctgaaccg
     2101 tactgcagac aaaatctgct tttagaggca agcggatttc tgacaaagta actgatcctt
     2161 tggatggcat aaattcactt tggggactag ccttattctt cctctgaggt ccttcgttct
     2221 tcaatttatt caattcatca atcaaaagtg ttctcttccc agttgcaatt agaagaagtc
     2281 tttctgcttc agcttcttct agggaccctt ttccatgttc ttcatcaaca cagcagttaa
     2341 gagcctggct agcttgatag atcactgtct gttgcatatt tatttcgtta ttgagttcct
     2401 gcattttctg tttgatatta acttgacaag gaaaggcatt atttttttca tccagttttg
     2461 aagtaacatc ttccttccga acaatcacct gctttattga tggacgttct gtttctttga
     2521 atctttgaga tctatatgca tcaatgctgt aaagaagatc acgatcttca gaaccaaggc
     2581 tatcaccaga ttcagctcga gggacacgag ttctcgaagc ttgagctcga gatctgagtc
     2641 cggacttgta cagctcgtcc atgccgccgg tggagtggcg gccctcggcg cgttcgtact
     2701 gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc caacttgatg ttgacgttgt
     2761 aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt ggtcttgacc tcagcgtcgt
     2821 agtggccgcc gtccttcagc ttcagcctct gcttgatctc gcccttcagg gcgccgtcct
     2881 cggggtacat ccgctcggag gaggcctccc agcccatggt cttcttctgc attacggggc
     2941 cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta gatgaactcg ccgtcctgca
     3001 gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc gaagttcatc acgcgctccc
     3061 acttgaagcc ctcggggaag gacagcttca agtagtcggg gatgtcggcg gggtgcttca
     3121 cgtaggcctt ggagccgtac atgaactgag gggacaggat gtcccaggcg aagggcaggg
     3181 ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc ctcgtagggg cggccctcgc
     3241 cctcgccctc gatctcgaac tcgtggccgt tcacggagcc ctccatgtgc accttgaagc
     3301 gcatgaactc cttgatgatg gccatgttat cctcctcgtc catgctcacc atggtggcga
     3361 ccggtagcgc tagcggatct gacggttcac taaaccagct ctgcttatat agacctccca
     3421 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     3481 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     3541 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     3601 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     3661 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     3721 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3781 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3841 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3901 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3961 actaatgcat ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4801 caatctaaag tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg
     4861 ctgcgagccc tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc
     4921 gggcgtattg gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc
     4981 gaaccccgcg tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg
     5041 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     5101 tagggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     5161 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     5221 gtgatggcag gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     5281 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     5341 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     5401 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     5461 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
     5521 atcctcgccg tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc
     5581 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
     5641 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     5701 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     5761 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     5821 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     5881 ctcgtcttgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     5941 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     6001 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     6061 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatcg atctttgcaa aagcctaggc
     6121 ctccaaaaaa g
//
