LOCUS       sequence-363413-        8859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector encoding myocardin-related transcription
            factor A tagged with mTagBFP2..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  AUTHORS   Zhang JZ, Nguyen AH, Miyamoto S, Heller Brown J, McCulloch AD, Zhang
            J
  TITLE     Histamine-induced biphasic activation of RhoA allows for persistent 
            RhoA signaling.
  JOURNAL   PLoS Biol. 2020 Sep 3;18(9):e3000866. doi: 
            10.1371/journal.pbio.3000866. eCollection 2020 Sep.
  PUBMED    32881857
REFERENCE   2  (bases 1 to 8859)
  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: "PLoS 
            Biol."; date: "2020-09-3"; volume: "18(9):e3000866. doi"; pages: " 
            10.1371/journal.pbio.3000866. eCollection 2020 Sep"
COMMENT     Sequence Label: pcDNA3.1(+)-mTagBFP2-MRTF-A
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             925..1623
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             complement(1978..1989)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4472..4489)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4478..4702
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4748..5175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4835..4854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5044..5065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5184..5204)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5349..5483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5410..5429
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5564..6358
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5618..5637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6228..6247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6532..6653
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     6623..6642
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6702..6718)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6702..6718)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6715..6737)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6726..6742
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6750..6756,6757..6774,6775..6780))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6795..6816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6933..6950)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7104..7692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7184..7203)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7863..8654,8655..8723))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8486..8505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8724..8828)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttgggc caccatgagc gagctgatta aggagaacat gcacatgaag
      961 ctgtacatgg agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc
     1021 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
     1081 ttcgccttcg acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac
     1141 acccagggca tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga
     1201 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     1261 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg
     1321 atgcagaaga aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc
     1381 ggcctggaag gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca
     1441 aacgccaaga ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc
     1501 tactatgtgg actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag
     1561 cagcacgagg tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt
     1621 aatggaggta ctggtggaag tatgccgcct ttgaaaagtc cagccgcatt tcatgagcag
     1681 agaaggagct tggagcgggc caggacagag gactatctca aacggaagat tcgttcccgg
     1741 ccggagagat cggagctggt caggatgcac attttggaag agacctcggc tgagccatcc
     1801 ctccaggcca agcagctgaa gctgaagaga gccagactag ccgatgacct caatgagaag
     1861 attgcacaga ggcctggccc catggagctg gtggagaaga acatccttcc tgttgagtcc
     1921 agcctgaagg aagccatcat tgtgggccag gtgaactatc ccaaagtagc agacagctct
     1981 tccttcgatg aggacagcag cgatgcctta tcccccgagc agcctgccag ccatgagtcc
     2041 cagggttctg tgccgtcacc cctggaggcc cgagtcagcg aaccactgct cagtgccacc
     2101 tctgcatccc ccacccaggt tgtgtctcaa cttccgatgg gccgggattc cagagaaatg
     2161 cttttcctgg cagagcagcc tcctctgcct cccccacctc tgctgcctcc cagcctcacc
     2221 aatggaacca ctatccccac tgccaagtcc acccccacac tcattaagca aagccaaccc
     2281 aagtctgcca gtgagaagtc acagcgcagc aagaaggcca aggagctgaa gccaaaggtg
     2341 aagaagctca agtaccacca gtacatcccc ccggaccaga agcaggacag gggggcaccc
     2401 cccatggact catcctacgc caagatcctg cagcagcagc agctcttcct ccagctgcag
     2461 atcctcaacc agcagcagca gcagcaccac aactaccagg ccatcctgcc tgccccgcca
     2521 aagtcagcag gcgaggccct gggaagcagc gggacccccc cagtacgcag cctctccact
     2581 accaatagca gctccagctc gggcgcccct gggccctgtg ggctggcacg tcagaacagc
     2641 acctcactga ctggcaagcc gggagccctg ccggccaacc tggacgacat gaaggtggca
     2701 gagctgaagc aggagctgaa gttgcgatca ctgcctgtct cgggcaccaa aactgagctg
     2761 attgagcgcc ttcgagccta tcaagaccaa atcagccctg tgccaggagc ccccaaggcc
     2821 cctgccgcca cctctatcct gcacaaggct ggcgaggtgg tggtagcctt cccagcggcc
     2881 cggctgagca cggggccagc cctggtggca gcaggcctgg ctccagctga ggtggtggtg
     2941 gccacggtgg ccagcagtgg ggtggtgaag tttggcagca cgggctccac gccccccgtg
     3001 tctcccaccc cctcggagcg ctcactgctc agcacgggcg atgaaaactc cacccccggg
     3061 gacacctttg gtgagatggt gacatcacct ctgacgcagc tgaccctgca ggcctcgcca
     3121 ctgcagatcc tcgtgaagga ggagggcccc cgggccgggt cctgttgcct gagccctggg
     3181 gggcgggcgg agctagaggg gcgcgacaag gaccagatgc tgcaggagaa agacaagcag
     3241 atcgaggcgc tgacgcgcat gctccggcag aagcagcagc tggtggagcg gctcaagctg
     3301 cagctggagc aggagaagcg agcccagcag cccgcccccg cccccgcccc cctcggcacc
     3361 cccgtgaagc aggagagcag cttctccagc tgccagctga gccagcagcc cctgggcccc
     3421 gctcacccat tcaaccccag cctggcggcc ccagccacca accacataga cccttgtgct
     3481 gtggccccgg ggcccccgtc cgtggtggtg aagcaggaag ccttgcagcc tgagcccgag
     3541 ccggtccccg ccccccagtt gcttctgggg cctcagggcc ccagcctcat caagggggtt
     3601 gcacctccca ccctcatcac cgactccaca gggacccacc ttgtcctcac cgtgaccaat
     3661 aagaatgcag acagccctgg cctgtccagt gggagccccc agcagccctc gtcccagcct
     3721 ggctctccag cgcctgcccc ctctgcccag atggacctgg agcacccact gcagcccctc
     3781 tttggggccc ccacttctct gctgaagaag gaaccacctg gctatgagga agccatgagc
     3841 cagcagccca aacagcagga aaatggttcc tcaagccagc agatggacga cctgtttgac
     3901 attctcattc agagcggaga aatttcagca gatttcaaag agccgccatc cctgccaggg
     3961 aaggagaagc catccccgaa gacagtctgt gggtcccccc tggcagcaca gccatcacct
     4021 tctgctgagc tcccccaggc tgccccacct cctccaggct caccctccct ccctggacgc
     4081 ctggaggact tcctggagag cagcacgggg ctgcccctgc tgaccagtgg gcatgacggg
     4141 ccagagcccc tttccctcat tgacgacctc catagccaga tgctgagcag cactgccatc
     4201 ctggaccacc ccccgtcacc catggacacc tcggaattgc actttgttcc tgagcccagc
     4261 agcaccatgg gcctggacct ggctgatggc cacctggaca gcatggactg gctggagctg
     4321 tcgtcaggtg gtcccgtgct gagcctagcc cccctcagca ccacagcccc cagcctcttc
     4381 tccacagact tcctcgatgg ccatgatttg cagctgcact gggattcctg cttgtagtag
     4441 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4501 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4561 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4621 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4681 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4741 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4801 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4861 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4921 cgatttagtg ctttacggca cctcgacccc aaaaacttga ttagggtgat ggttcacgta
     4981 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5041 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5101 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5161 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5221 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5281 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5341 aaccatagtc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5401 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     5461 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttatatatc
     5521 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5581 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5641 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5701 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5761 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5821 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5881 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5941 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     6001 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     6061 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     6121 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     6181 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     6241 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     6301 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     6361 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     6421 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     6481 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     6541 attgcagctt ataatggtta caaataaagc aatagaatca caaatttcac aaataaagca
     6601 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     6661 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6721 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6781 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6841 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6901 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6961 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7021 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     7081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     7141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     7201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     7261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     7321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     7381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     7441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     7501 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     7561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     7621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     7681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7921 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7981 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8041 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8101 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8161 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8221 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8281 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8341 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8401 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     8461 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     8521 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     8581 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     8641 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     8701 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8761 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8821 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
