LOCUS       sequence-197860-        6140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Membrane marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6140)
  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 6140)
  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: mCherry-KRasCT
FEATURES             Location/Qualifiers
     source          1..6140
                     /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"
     primer_bind     complement(3390..3408)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3658..3677
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(4059..4076)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4065..4289
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4335..4763
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4422..4441)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4632..4653
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4772..4792)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4777..5106
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4957..5092
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5019..5038
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5173..5967
                     /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(5227..5246)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5837..5856
                     /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 gcttatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     3301 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     3361 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     3421 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     3481 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     3541 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     3601 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     3661 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     3721 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     3781 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     3841 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     3901 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagcttacaa ggaattcgga
     3961 agcggaagcg ggagtgggga gcaaaaagaa gaaaaagaag tcaaagacaa agtgtgtaat
     4021 tatgtagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     4081 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4141 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4201 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4261 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     4321 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4381 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4681 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4741 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     4801 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4861 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4921 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4981 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     5041 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     5101 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     5161 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5221 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5281 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5341 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5401 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5461 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5521 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5581 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5641 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5701 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5761 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5821 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     5881 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     5941 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6001 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6061 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6121 tggagttctt cgcccacccc
//
