LOCUS       sequence-197780-        7625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RhoA mediated cell migration.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7625)
  AUTHORS   O'Neill PR, Castillo-Badillo JA, Meshik X, Kalyanaraman V, Melgarejo
            K, Gautam N
  TITLE     Membrane Flow Drives an Adhesion-Independent Amoeboid Cell Migration
            Mode.
  JOURNAL   Dev Cell. 2018 Jun 15. pii: S1534-5807(18)30422-2. doi: 
            10.1016/j.devcel.2018.05.029.
  PUBMED    29937389
REFERENCE   2  (bases 1 to 7625)
  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: "Dev Cell. 
            2018 Jun 15. pii: S1534-5807(18)30422-2. doi: 
            10.1016/j.devcel.2018.05.029."
COMMENT     Sequence Label: LARG mCherry sspB in pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(38..57)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     92..111
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(171..187)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(171..187)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(184..206)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    195..211
                     /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(219..225,226..243,244..249))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    264..285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(402..419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(573..1161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(653..672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1332..2123,2124..2192))
                     /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     1955..1974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2193..2297)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2372..2391)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2563..2942
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2943..3146
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3097..3117
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3191..3210
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3191..3209
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4394..5101
                     /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(4539..4557)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4807..4826
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(5177..5383,5384..5386,5387..5506)
                     /codon_start=1
                     /product="SspB from Haemophilus influenzae, with the 
                     mutations Y11K and A15E to disrupt the natural dimerization
                     of the protein and with the mutation R74Q to reduce the 
                     affinity for iLID"
                     /label=SspB micro
                     /note="dimerizes with iLID at moderate affinity to create 
                     iLID micro (Guntas et al., 2015)"
                     /translation="SSPKRPKLLREYYDWLVDNSFTPYLVVDATYLGVNVPVEYVKDGQ
                     IVLNLSASATGNLQLTNDFIQFNAQFKGVSRELYIPMGAALAIYARENGDGVMFEPEEI
                     YDELNI"
     primer_bind     complement(5544..5561)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5550..5774
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5820..6248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5907..5926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6117..6138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6257..6277)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6262..6591
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6442..6577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6504..6523
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6658..7452
                     /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(6712..6731)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7322..7341
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
      181 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      241 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      301 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      361 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      421 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      481 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      541 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      601 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      661 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      721 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      781 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      841 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      901 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      961 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1021 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1081 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1141 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1201 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1261 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1321 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1381 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1441 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     1501 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1561 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1621 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1681 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1741 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1801 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1861 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1921 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1981 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     2041 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     2101 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2161 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2221 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2281 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     2341 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2401 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     2461 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     2521 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     2581 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     2641 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     2701 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     2761 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     2821 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     2881 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     2941 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     3001 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     3061 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     3121 ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact gcttactggc
     3181 ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt ttaaacttaa
     3241 gcttggtacc atgcccaact ggcagcagct tgttagtcga gaagtgttac tgggactaaa
     3301 accttgtgaa atcaaaagac aggaagtgat taatgaattg ttctacactg aaagagctca
     3361 tgttcgaaca ctgaaggttc ttgatcaagt gttctatcag cgagtatcca gagaaggaat
     3421 tctgtcaccc tcagagctac ggaaaatttt ttcaaacttg gaagatattc ttcaacttca
     3481 tattggattg aatgaacaaa tgaaggctgt tcgaaagaga aatgagacct ctgttatcga
     3541 tcagattggg gaagatttgc tgacatggtt cagcggacca ggagaggaga aattgaaaca
     3601 tgctgctgct accttttgca gtaaccaacc tttcgccctg gaaatgatca aatctcgtca
     3661 gaaaaaggat tctcgatttc agacttttgt gcaagatgct gaaagtaatc cactgtgtcg
     3721 tcgtcttcaa ctgaaggata ttattcccac tcaaatgcaa aggcttacta agtacccact
     3781 tctgttggat aatattgcca aatacacaga atggccaaca gaaagggaga aggtgaagaa
     3841 agctgcagat cactgtcgtc agatcttaaa ttttgtaaat caggctgtca aggaggcaga
     3901 aaacaagcag cgcctagaag attatcagcg tcgccttgat acctccagcc tgaagttgtc
     3961 agagtaccca aatgttgaag agctcaggaa tttggattta acaaaaagga agatgattca
     4021 tgaagggcca ttggtttgga aggtgaatag agataaaact attgatttat acacgttgct
     4081 gctggaagac attcttgtat tgttacaaaa gcaggatgat agactggttt taaggtgtca
     4141 tagtaagatt ctggcatcta cagctgatag caaacacacg tttagccctg tcattaagtt
     4201 gagtacagtg ttggttcgac aagtggcaac agataacaaa gctttattcg tcatttccat
     4261 gtcagacaat ggcgctcaga tttatgaact ggtggcacag acagtttctg aaaagactgt
     4321 ctggcaggac ctaatctgtc ggatggctgc atcagtgaag gagcaatccg gatccggtag
     4381 tggaagtggt tccatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     4441 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     4501 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     4561 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     4621 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     4681 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     4741 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     4801 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     4861 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     4921 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     4981 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     5041 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     5101 gtccggactc agatctcgag ctcaagcttc gaacgaattc ggcattgatc tgagcggcct
     5161 gaccctgcag gaattcagct ccccgaaacg ccctaagctg ctgcgtgaat attacgattg
     5221 gctggttgat aacagcttta ccccatatct ggtggtggat gccacatacc tgggcgtgaa
     5281 cgtgcccgtg gagtatgtga aagacggtca gatcgtgctg aatctgtctg caagtgcgac
     5341 cggcaacctg caactgacaa atgattttat ccagttcaac gcccagttta agggcgtgtc
     5401 tcgtgaactg tatatcccga tgggtgccgc tctggccatt tacgctcgcg agaacggcga
     5461 tggtgtgatg ttcgaaccag aagaaatcta tgacgagctg aatattggtt aatctagagg
     5521 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5581 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5641 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5701 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     5761 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5821 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5881 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5941 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     6001 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     6061 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     6121 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     6181 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     6241 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     6301 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     6361 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     6421 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     6481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     6541 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     6601 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     6661 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     6721 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6781 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6841 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6901 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6961 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     7021 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     7081 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     7141 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     7201 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     7261 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     7321 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     7381 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     7441 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     7501 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     7561 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     7621 acccc
//
