LOCUS       sequence-197861-        6801 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Localization and visualization of trafficking protein Rab4 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  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 6801)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: YFP Rab4
FEATURES             Location/Qualifiers
     source          1..6801
                     /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             3233..3946
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             3233..3946
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3263..3291)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3886..3906
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     protein_bind    4007..4031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4720..4737)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4726..4950
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4996..5424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5083..5102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5293..5314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5433..5453)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5438..5767
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5618..5753
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5680..5699
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5834..6628
                     /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(5888..5907)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6498..6517
                     /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 tggctagcta gcatgagtaa
     3241 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     3301 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     3361 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
     3421 tttcggttat ggtcttatgt gctttgcaag atacccagat catatgaaac agcatgactt
     3481 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     3541 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     3601 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggagta
     3661 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
     3721 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     3781 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccta
     3841 tcaatctaag ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     3901 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaaggat ctcgagctca
     3961 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tcatcaacaa gtttgtacaa
     4021 aaaagcaggc ttaatgtcgg aaacatacga ttacctgttc aagttcctga taattggcag
     4081 tgctggcagc ggtaagagtt gtctgttgca tcacttcatc gagagcaagt tcaaagacga
     4141 cagttcgcac accatcggag tggagttcgg ctcgcggatt gtgaacgtgg gcggcaagtc
     4201 ggtaaagcta cagatatggg acacagccgg tcaggagaga ttccgatcgg tgacaagatc
     4261 gtactatcgc ggagcagccg gagctctcct ggtctacgat gccacttcgc gggactcttt
     4321 caatgctttg acaaactggc tgaatgatgc tcgcactctg gccagcccaa acattgtgat
     4381 ccttttggtg ggcaataaaa aggacttgga ggaggcgcgt gatgtgacct ttctggaagc
     4441 aagcaccttt gcccaagaga acgagcttat cttcctggag acgagcgcca agacggggga
     4501 gaatgttgaa gaggcctttc tcaagtgctc aaagaccata ctggccaaaa ttgagaccgg
     4561 agagctggat cccgagcgca tcggcagcgg tattcagtac ggcggagctg ctctccgtaa
     4621 cttacagaca cgacagcgca gcatcaataa gccggattgc acctgtcgcg tttaagcggc
     4681 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4741 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4801 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4861 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     4921 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     4981 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5041 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5101 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5161 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5221 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5281 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5341 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     5401 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     5461 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5521 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     5581 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     5641 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     5701 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     5761 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     5821 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5881 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5941 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6001 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6061 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     6121 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     6181 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     6241 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     6301 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     6361 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     6421 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     6481 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     6541 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6601 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6661 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6721 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6781 ctggagttct tcgcccaccc c
//
