LOCUS       Exported                7097 bp ds-DNA     circular SYN 14-SEP-2022
DEFINITION  Express pmCherry-IL2-AH-2A-RhoA Q63L .
ACCESSION   .
VERSION     .
KEYWORDS    IL2-AH-2A-Cherry-RhoAQ63L in pmCherryN1
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 Sep 14, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7097
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        73..376
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        377..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     531..551
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2408..2461
                     /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             2480..3187
                     /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(2625..2643)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2893..2912
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3194..3775
                     /codon_start=1
                     /product="constitutively active mutant of human Rho family 
                     small GTPase RhoA"
                     /label=RhoA (Q63L)
                     /translation="MAAIRKKLVIVGDGACGKTCLLIVFSKDQFPEVYVPTVFENYVAD
                     IEVDGKQVELALWDTAGLEDYDRLRPLSYPDTDVILMCFSIDSPDSLENIPEKWTPEVK
                     HFCPNVPIILVGNKKDLRNDEHTRRELAKMKQEPVKPEEGRDMANRIGAFGYMECSAKT
                     KDGVREVFEMATRAALQARRGKKKSGCLVL"
     polyA_signal    3897..4018
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3934..3953)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3988..4007
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4025..4480)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4162..4183)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4374..4393
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4507..4611
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4613..4970
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4636..4656)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4821..4956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4883..4902
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5005..5799
                     /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(5059..5078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5669..5688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5987..6006)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6031..6078
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6407..6995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6896..6915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
       61 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      121 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      181 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      241 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      301 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      361 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      421 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      481 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      541 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      601 ccgctagatg gattcatacc tgctgatgtg gggactgctc acgttcatca tggtgcctgg
      661 ctgccaggca gagctctgtg acgatgaccc gccagagatc ccacacgcca cattcaaagc
      721 catggcctac aaggaaggaa ccatgttgaa ctgtgaatgc aagagaggtt tccgcagaat
      781 aaaaagcggg tcactctata tgctctgtac aggaaactct agccactcgt cctgggacaa
      841 ccaatgtcaa tgcacaagct ctgccactcg gaacacaacg aaacaagtga cacctcaacc
      901 tgaagaacag aaagaaagga aaaccacaga aatgcaaagt ccaatgcagc cagtggacca
      961 agcgagcctt ccaggtcact gcagggaacc tccaccatgg gaaaatgaag ccacagagag
     1021 aatttatcat ttcgtggtgg ggcagatggt ttattatcag tgcgtccagg gatacagggc
     1081 tctacacaga ggtcctgctg agagcgtctg caaaatgacc cacgggaaga caaggtggac
     1141 ccagccccag ctcatatgca caggtgaaat ggagaccagt cagtttccag gtgaagagaa
     1201 gcctcaggca agccccgaag gccgtcctga gagtgagact tcctgcctcg tcacaacaac
     1261 agattttcaa atacagacag aaatggctgc aaccatggag acgtccatat ttacaacaga
     1321 gtaccaggta gcagtggccg gctgtgtttt cctgctgatc agcgtcctcc tcctgagtgg
     1381 gctcacctgg aagggaaaat tccaaagaac tcgtgtccct cgagctgaat ctggtgatag
     1441 ccttggttct gaagatcgtg atcttcttta cagcattgat gcatatagat ctcaaagatt
     1501 caaagaaaca gaacgtccat caataaagca ggtgattgtt cggaaggaag atgttacttc
     1561 aaaactggat gaaaaaaata atgcctttcc ttgtcaagtt aatatcaaac agaaaatgca
     1621 ggaactcaat aacgaaataa atatgcaaca gacagtgatc tatcaagcta gccaggctct
     1681 taactgctgt gttgatgaag aacatggaaa agggtcccta gaagaagctg aagcagaaag
     1741 acttcttcta attgcaactg ggaagagaac acttttgatt gatgaattga ataaattgaa
     1801 gaacgaagga cctcagagga agaataaggc tagtccccaa agtgaattta tgccatccaa
     1861 aggatcagtt actttgtcag aaatccgctt gcctctaaaa gcagattttg tctgcagtac
     1921 ggttcagaaa ccagatgcag caaattacta ttacttaatt atactaaaag caggagctga
     1981 aaatatggta gccacaccat tagcaagtac ttcaaactct cttaacggtg atgctctgac
     2041 attcactact acatttactc tgcaagatgt atccaatgac tttgaaataa atattgaagt
     2101 ttacagcttg gtgcaaaaga aagatccctc aggccttgat aagaagaaaa aaacatccaa
     2161 gtccaaggct attactccaa agcgactcct cacatctata accacaaaaa gcaacattca
     2221 ttcttcagtc atggccagtc caggaggtct tagtgctgtg cgaaccagca acttcgccct
     2281 tgttggatct tacacattat cattgtcttc agtaggaaat actaagtttg ttctggacaa
     2341 ggtccccttt ttatcttctt tggaaggtca tatttattta aaaataaaat gtcaagtgcg
     2401 accggtcgag ggcaggggaa gcctgctcac ctgcggcgac gtggaggaaa accctggccc
     2461 cgccccagga tccaagctta tggtgagcaa gggcgaggag gataacatgg ccatcatcaa
     2521 ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat
     2581 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac
     2641 caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc
     2701 caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga
     2761 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca
     2821 ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt
     2881 cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct cctccgagcg
     2941 gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga
     3001 cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct
     3061 gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac
     3121 catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct
     3181 gtacaaggaa ttcatggctg ccatccggaa gaaactggtg attgttggtg atggagcctg
     3241 tggaaagaca tgcttgctca tagtcttcag caaggaccag ttcccagagg tgtatgtgcc
     3301 cacagtgttt gagaactatg tggcagatat cgaggtggat ggaaagcagg tagagttggc
     3361 tttgtgggac acagctgggc tggaagatta tgatcgcctg aggcccctct cctacccaga
     3421 taccgatgtt atactgatgt gtttttccat cgacagccct gatagtttag aaaacatccc
     3481 agaaaagtgg accccagaag tcaagcattt ctgtcccaac gtgcccatca tcctggttgg
     3541 gaataagaag gatcttcgga atgatgagca cacaaggcgg gagctagcca agatgaagca
     3601 ggagccggtg aaacctgaag aaggcagaga tatggcaaac aggattggcg cttttgggta
     3661 catggagtgt tcagcaaaga ccaaagatgg agtgagagag gtttttgaaa tggctacgag
     3721 agctgctctg caagctagac gtgggaagaa aaaatctggt tgccttgtct tgtgagcggc
     3781 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     3841 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     3901 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3961 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag
     4021 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4081 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4141 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4201 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4261 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4321 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4381 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     4441 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga
     4501 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4561 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg
     4621 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4681 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4741 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4801 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4861 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4921 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa
     4981 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5041 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5101 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5161 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg
     5221 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5281 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5341 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5401 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5461 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5521 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5581 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5641 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5701 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5761 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5821 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5881 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5941 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg
     6001 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     6061 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata
     6121 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc
     6181 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca
     6241 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6301 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6361 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6421 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6481 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6541 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6601 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6661 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6721 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6781 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6841 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6901 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6961 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7021 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7081 ctgtggataa ccgtatt
//
