LOCUS       sequence-429702-        4279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Volume marker / Control sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4279)
  AUTHORS   Nanda S, Calderon A, Sachan A, Duong TT, Koch J, Xin X, 
            Solouk-Stahlberg D, Wu YW, Nalbant P, Dehmelt L
  TITLE     Rho GTPase activity crosstalk mediated by Arhgef11 and Arhgef12 
            coordinates cell protrusion-retraction cycles.
  JOURNAL   Nat Commun. 2023 Dec 15;14(1):8356. doi: 10.1038/s41467-023-43875-y.
  PUBMED    38102112
REFERENCE   2  (bases 1 to 4279)
  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: "Nat 
            Commun."; date: "2023-12-15"; volume: "14(1):8356. doi"; pages: " 
            10.1038/s41467-023-43875-y"
COMMENT     Sequence Label: delCMV-mCitrine
FEATURES             Location/Qualifiers
     source          1..4279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..30
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    174..295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(185..204)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     239..258
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(418..1131,1132..1134,1135..1137))
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(463..484)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     812..831
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1071..1092
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1145..1225
                     /label=MCS
                     /note="multiple cloning site"
     rep_origin      complement(1476..2064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1556..1575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2393..2440
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2465..2484
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2672..3466)
                     /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(2672..3466)
                     /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(2783..2802)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3393..3412
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3501..3858)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3515..3650
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3667..3858
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     3815..3835
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(3860..3964)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(3991..4279,1..167)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4078..4097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
       61 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      121 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttacg ccttaagata
      181 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
      241 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
      301 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
      361 caagtaaaac ctctacaaat gtggtatggc tgattatgat ctagagtcgc ggccgcttta
      421 cttgtacagc tcgtccatgc cgagagtgat cccggcggcg gtcacgaact ccagcaggac
      481 catgtgatcg cgcttctcgt tggggtcttt gctcagcttg gactggtagc tcaggtagtg
      541 gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc
      601 gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg aagttcacct tgatgccgtt
      661 cttctgcttg tcggccatga tatagacgtt gtggctgttg tagttgtact ccagcttgtg
      721 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
      781 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa
      841 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
      901 catgtggtcg gggtagcggg cgaagcacat caggccgtag ccgaaggtgg tcacgagggt
      961 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta
     1021 ggtggcatcg ccctcgccct cgccggacac gctgaacttg tggccgttta cgtcgccgtc
     1081 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcaccatggt
     1141 ggcgaccggt ggatcccggg cccgcggtac cgtcgactgc agaattcgaa gcttgagctc
     1201 gagatctgag tccggtagcg ctagcggatc tgacggttca ctaaaccagc tctgcttata
     1261 tagacctccc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     1321 tatgaactaa tgaccccgta attgattact attaataact aatgcatggc ggtaatacgg
     1381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2221 cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg gccttcaccc
     2281 gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc ccaatggggt
     2341 ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac
     2401 gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg ggttccttcc
     2461 ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa gaactccagc
     2521 atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat tccgaagccc
     2581 aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt gggcgtcgct
     2641 tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga
     2701 aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag cggtcagccc
     2761 attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc tgatagcggt
     2821 ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt tccaccatga
     2881 tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg ggcatgctcg
     2941 ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg tccagatcat
     3001 cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt
     3061 ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca
     3121 tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc cccggcactt
     3181 cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag
     3241 gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg
     3301 caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca
     3361 cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat agcctctcca
     3421 cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga aacgatcctc
     3481 atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
     3541 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
     3601 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
     3661 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     3721 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
     3781 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacagc
     3841 tggttctttc cgcctcagga ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3961 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     4021 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4081 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4141 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4201 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4261 tgacgttgga gtccacgtt
//
