LOCUS       sequence-372005-        6959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express pmCherry-IL2-AH-deltaC2-2A-RhoA Q63L .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6959)
  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 6959)
  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-AHDC2-2A-CherryRhoAQ63L in pmCherryN1
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2266..2319
                     /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             2338..3045
                     /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(2483..2501)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2751..2770
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(3052..3237,3238..3240,3241..3633)
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase RhoA"
                     /label=RhoA (Q63L)
                     /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    3755..3876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3792..3811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3846..3865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3883..4338)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4020..4041)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4232..4251
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4365..4469
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4471..4828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4471..4662
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4494..4514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4679..4814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4863..5657
                     /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             4863..5657
                     /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(4917..4936)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5527..5546
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5845..5864)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5889..5936
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6265..6853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6754..6773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagatggatt catacctgct gatgtgggga ctgctcacgt tcatcatggt gcctggctgc
      661 caggcagagc tctgtgacga tgacccgcca gagatcccac acgccacatt caaagccatg
      721 gcctacaagg aaggaaccat gttgaactgt gaatgcaaga gaggtttccg cagaataaaa
      781 agcgggtcac tctatatgct ctgtacagga aactctagcc actcgtcctg ggacaaccaa
      841 tgtcaatgca caagctctgc cactcggaac acaacgaaac aagtgacacc tcaacctgaa
      901 gaacagaaag aaaggaaaac cacagaaatg caaagtccaa tgcagccagt ggaccaagcg
      961 agccttccag gtcactgcag ggaacctcca ccatgggaaa atgaagccac agagagaatt
     1021 tatcatttcg tggtggggca gatggtttat tatcagtgcg tccagggata cagggctcta
     1081 cacagaggtc ctgctgagag cgtctgcaaa atgacccacg ggaagacaag gtggacccag
     1141 ccccagctca tatgcacagg tgaaatggag accagtcagt ttccaggtga agagaagcct
     1201 caggcaagcc ccgaaggccg tcctgagagt gagacttcct gcctcgtcac aacaacagat
     1261 tttcaaatac agacagaaat ggctgcaacc atggagacgt ccatatttac aacagagtac
     1321 caggtagcag tggccggctg tgttttcctg ctgatcagcg tcctcctcct gagtgggctc
     1381 acctggaagg gaaaattcca aagaactcgt gtccctcgag ctgaatctgg tgatagcctt
     1441 ggttctgaag atcgtgatct tctttacagc attgatgcat atagatctca aagattcaaa
     1501 gaaacagaac gtccatcaat aaagcaggtg attgttcgga aggaagatgt tacttcaaaa
     1561 ctggatgaaa aaaataatgc ctttccttgt caagttaata tcaaacagaa aatgcaggaa
     1621 ctcaataacg aaataaatat gcaacagaca gtgatctatc aagctagcca ggctcttaac
     1681 tgctgtgttg atgaagaaca tggaaaaggg tccctagaag aagctgaagc agaaagactt
     1741 cttctaattg caactgggaa gagaacactt ttgattgatg aattgaataa attgaagaac
     1801 gaaggacctc agaggaagaa taaggctagt ccccaaagtg aatttatgcc atccaaagga
     1861 tcagttactt tgtcagaaat ccgcttgcct ctaaaagcag attttgtctg cagtacggtt
     1921 cagaaaccag atgcagcaaa ttactattac ttaattatac taaaagcagg agctgaaaat
     1981 atggtagcca caccattagc aagtacttca aactctctta acggtgatgc tctgacattc
     2041 actactacat ttactctgca agatgtatcc aatgactttg aaataaatat tgaagtttac
     2101 agcttggtgc aagctgtgcg aaccagcaac ttcgcccttg ttggatctta cacattatca
     2161 ttgtcttcag taggaaatac taagtttgtt ctggacaagg tccccttttt atcttctttg
     2221 gaaggtcata tttatttaaa aataaaatgt caagtgcgac cggtcgaggg caggggaagc
     2281 ctgctcacct gcggcgacgt ggaggaaaac cctggccccg ccccaggatc caagcttatg
     2341 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
     2401 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2461 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2521 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
     2581 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2641 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
     2701 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     2761 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
     2821 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
     2881 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
     2941 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
     3001 gccgagggcc gccactccac cggcggcatg gacgagctgt acaaggaatt catggctgcc
     3061 atccggaaga aactggtgat tgttggtgat ggagcctgtg gaaagacatg cttgctcata
     3121 gtcttcagca aggaccagtt cccagaggtg tatgtgccca cagtgtttga gaactatgtg
     3181 gcagatatcg aggtggatgg aaagcaggta gagttggctt tgtgggacac agctgggctg
     3241 gaagattatg atcgcctgag gcccctctcc tacccagata ccgatgttat actgatgtgt
     3301 ttttccatcg acagccctga tagtttagaa aacatcccag aaaagtggac cccagaagtc
     3361 aagcatttct gtcccaacgt gcccatcatc ctggttggga ataagaagga tcttcggaat
     3421 gatgagcaca caaggcggga gctagccaag atgaagcagg agccggtgaa acctgaagaa
     3481 ggcagagata tggcaaacag gattggcgct tttgggtaca tggagtgttc agcaaagacc
     3541 aaagatggag tgagagaggt ttttgaaatg gctacgagag ctgctctgca agctagacgt
     3601 gggaagaaaa aatctggttg ccttgtcttg tgagcggccg cgactctaga tcataatcag
     3661 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3721 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3781 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3841 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3901 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3961 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     4021 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4081 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4141 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4201 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4261 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4321 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4381 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4441 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4501 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4561 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4621 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4681 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4741 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4801 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4861 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4921 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4981 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5041 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5101 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5161 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5221 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5281 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5341 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5401 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5461 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5521 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5581 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5641 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5701 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5761 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5821 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5881 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5941 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     6001 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6061 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6121 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6181 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6241 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6301 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6361 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6421 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6481 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6541 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6601 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6661 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6721 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6781 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6841 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6901 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccg
//
