LOCUS       sequence-372033-        7097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express pmCherry-IL2-AH A740D, E758K-2A-RhoA Q63L .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
  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 7097)
  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: IL2-AHA740D, E758K-2A-Cherry-RhoAQ63L in pmChe...
FEATURES             Location/Qualifiers
     source          1..7097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        79..382
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        383..586
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     537..557
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2414..2467
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             2486..3193
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2631..2649)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2899..2918
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(3200..3385,3386..3388,3389..3781)
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase RhoA"
                     /label=RhoA (Q63L)
                     /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    3903..4024
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3940..3959)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3994..4013
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4031..4486)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4168..4189)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4380..4399
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4513..4617
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4619..4976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4619..4810
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4642..4662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4827..4962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5011..5805
                     /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             5011..5805
                     /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(5065..5084)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5675..5694
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5993..6012)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6037..6084
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6413..7001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6902..6921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
       61 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      121 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      181 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      241 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      301 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      361 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      421 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      481 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      541 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
      601 tcagatccgc tagatggatt catacctgct gatgtgggga ctgctcacgt tcatcatggt
      661 gcctggctgc caggcagagc tctgtgacga tgacccgcca gagatcccac acgccacatt
      721 caaagccatg gcctacaagg aaggaaccat gttgaactgt gaatgcaaga gaggtttccg
      781 cagaataaaa agcgggtcac tctatatgct ctgtacagga aactctagcc actcgtcctg
      841 ggacaaccaa tgtcaatgca caagctctgc cactcggaac acaacgaaac aagtgacacc
      901 tcaacctgaa gaacagaaag aaaggaaaac cacagaaatg caaagtccaa tgcagccagt
      961 ggaccaagcg agccttccag gtcactgcag ggaacctcca ccatgggaaa atgaagccac
     1021 agagagaatt tatcatttcg tggtggggca gatggtttat tatcagtgcg tccagggata
     1081 cagggctcta cacagaggtc ctgctgagag cgtctgcaaa atgacccacg ggaagacaag
     1141 gtggacccag ccccagctca tatgcacagg tgaaatggag accagtcagt ttccaggtga
     1201 agagaagcct caggcaagcc ccgaaggccg tcctgagagt gagacttcct gcctcgtcac
     1261 aacaacagat tttcaaatac agacagaaat ggctgcaacc atggagacgt ccatatttac
     1321 aacagagtac caggtagcag tggccggctg tgttttcctg ctgatcagcg tcctcctcct
     1381 gagtgggctc acctggaagg gaaaattcca aagaactcgt gtccctcgag ctgaatctgg
     1441 tgatagcctt ggttctgaag atcgtgatct tctttacagc attgatgcat atagatctca
     1501 aagattcaaa gaaacagaac gtccatcaat aaagcaggtg attgttcgga aggaagatgt
     1561 tacttcaaaa ctggatgaaa aaaataatgc ctttccttgt caagttaata tcaaacagaa
     1621 aatgcaggaa ctcaataacg aaataaatat gcaacagaca gtgatctatc aagctagcca
     1681 ggatcttaac tgctgtgttg atgaagaaca tggaaaaggg tccctagaag aagctaaagc
     1741 agaaagactt cttctaattg caactgggaa gagaacactt ttgattgatg aattgaataa
     1801 attgaagaac gaaggacctc agaggaagaa taaggctagt ccccaaagtg aatttatgcc
     1861 atccaaagga tcagttactt tgtcagaaat ccgcttgcct ctaaaagcag attttgtctg
     1921 cagtacggtt cagaaaccag atgcagcaaa ttactattac ttaattatac taaaagcagg
     1981 agctgaaaat atggtagcca caccattagc aagtacttca aactctctta acggtgatgc
     2041 tctgacattc actactacat ttactctgca agatgtatcc aatgactttg aaataaatat
     2101 tgaagtttac agcttggtgc aaaagaaaga tccctcaggc cttgataaga agaaaaaaac
     2161 atccaagtcc aaggctatta ctccaaagcg actcctcaca tctataacca caaaaagcaa
     2221 cattcattct tcagtcatgg ccagtccagg aggtcttagt gctgtgcgaa ccagcaactt
     2281 cgcccttgtt ggatcttaca cattatcatt gtcttcagta ggaaatacta agtttgttct
     2341 ggacaaggtc ccctttttat cttctttgga aggtcatatt tatttaaaaa taaaatgtca
     2401 agtgcgaccg gtcgagggca ggggaagcct gctcacctgc ggcgacgtgg aggaaaaccc
     2461 tggccccgcc ccaggatcca agcttatggt gagcaagggc gaggaggata acatggccat
     2521 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     2581 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa
     2641 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     2701 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt
     2761 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt
     2821 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac
     2881 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc
     2941 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct
     3001 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt
     3061 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga
     3121 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     3181 cgagctgtac aaggaattca tggctgccat ccggaagaaa ctggtgattg ttggtgatgg
     3241 agcctgtgga aagacatgct tgctcatagt cttcagcaag gaccagttcc cagaggtgta
     3301 tgtgcccaca gtgtttgaga actatgtggc agatatcgag gtggatggaa agcaggtaga
     3361 gttggctttg tgggacacag ctgggctgga agattatgat cgcctgaggc ccctctccta
     3421 cccagatacc gatgttatac tgatgtgttt ttccatcgac agccctgata gtttagaaaa
     3481 catcccagaa aagtggaccc cagaagtcaa gcatttctgt cccaacgtgc ccatcatcct
     3541 ggttgggaat aagaaggatc ttcggaatga tgagcacaca aggcgggagc tagccaagat
     3601 gaagcaggag ccggtgaaac ctgaagaagg cagagatatg gcaaacagga ttggcgcttt
     3661 tgggtacatg gagtgttcag caaagaccaa agatggagtg agagaggttt ttgaaatggc
     3721 tacgagagct gctctgcaag ctagacgtgg gaagaaaaaa tctggttgcc ttgtcttgtg
     3781 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     3841 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     3901 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3961 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4021 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     4081 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     4141 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     4201 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     4261 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4321 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4381 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4441 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     4501 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4561 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     4621 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4681 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4741 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4801 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4861 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     4921 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     4981 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     5041 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     5101 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     5161 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     5221 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5281 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5341 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5401 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5461 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5521 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5581 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5641 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5701 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5761 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5821 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5881 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5941 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     6001 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     6061 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     6121 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     6181 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     6241 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     6301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6541 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7081 ctgattctgt ggataac
//
