LOCUS       sequence-365843-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing paxillin variants in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  AUTHORS   Case LB, De Pasquale M, Henry L, Rosen MK
  TITLE     Synergistic phase separation of two pathways promotes integrin 
            clustering and nascent adhesion formation.
  JOURNAL   Elife. 2022 Jan 20;11. pii: 72588. doi: 10.7554/eLife.72588.
  PUBMED    35049497
REFERENCE   2  (bases 1 to 6423)
  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: "Elife."; 
            date: "2022-01-20"; volume: "11. pii: 72588. doi"; pages: " 
            10.7554/eLife.72588"
COMMENT     Sequence Label: GFP-Paxillin WT
FEATURES             Location/Qualifiers
     source          1..6423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      721..730
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(727..729,730..732,733..1443)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(772..793)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1033..1052)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1380..1401
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3325..3446
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3362..3381)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3416..3435
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3453..3908)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3590..3611)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3802..3821
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3935..4039
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4041..4398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4041..4232
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4064..4084)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4249..4384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4433..5227
                     /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             4433..5227
                     /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(4487..4506)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5097..5116
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5415..5434)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5459..5506
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5835..6423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6324..6343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      721 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      781 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      841 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      901 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      961 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1021 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1081 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1141 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1201 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1261 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1321 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1381 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1441 aagtccggac tcagatccgg aagcggcggt tccatggacg acctcgacgc cctgctggcg
     1501 gacttggagt ctaccacctc ccacatctcc aaacggcctg tgttcttgtc ggaggagacc
     1561 ccctactcat acccaactgg aaaccacaca taccaggaga ttgccgtgcc accccccgtc
     1621 cccccacccc cgtccagcga ggccctcaat ggcacaatcc ttgacccctt agaccagtgg
     1681 cagcccagcg gctcccgatt catccaccag cagcctcagt cctcatcacc tgtgtacggc
     1741 tccagtgcca aaacttccag tgtctccaac cctcaggaca gtgttggctc tccgtgctcc
     1801 cgagtgggtg aggaggagca cgtctacagc ttccccaaca agcagaaatc agctgagcct
     1861 tcacccaccg taatgagcac gtccctgggc agcaaccttt ctgaactcga ccgcctgctg
     1921 ctggaactga acgctgtaca gcataacccg ccaggcttcc ctgcagatga ggccaactca
     1981 agccccccgc ttcctggggc cctgagcccc ctctatggtg tcccagagac taacagcccc
     2041 ttgggaggca aagctgggcc cctgacgaaa gagaagccta agcggaatgg gggccggggc
     2101 ctggaggacg tgcggcccag tgtggagagt ctcttggatg aactggagag ctccgtgccc
     2161 agccccgtcc ctgccatcac tgtgaaccag ggcgagatga gcagcccgca gcgcgtcacc
     2221 tccacccaac agcagacacg catctcggcc tcctctgcca ccagggagct ggacgagctg
     2281 atggcttcgc tgtcggattt caagttcatg gcccagggga agacagggag cagctcaccc
     2341 cctggggggc ccccgaagcc cgggagccag ctggacagca tgctggggag cctgcagtct
     2401 gacctgaaca agctgggggt cgccacagtc gccaaaggag tctgcggggc ctgcaagaag
     2461 cccatcgccg ggcaggttgt gaccgccatg gggaagacgt ggcaccccga gcacttcgtc
     2521 tgcacccact gccaggagga gatcggttca cggaacttct tcgagcggga tggacagccc
     2581 tactgtgaaa aggactacca caacctcttc tccccgcgct gctactactg caacggcccc
     2641 atcctggata aagtggtgac agcccttgac cggacgtggc accctgaaca cttcttctgt
     2701 gcacagtgtg gagccttctt tggtcccgaa gggttccacg agaaggacgg caaggcctac
     2761 tgtcgcaagg actacttcga catgttcgca cccaagtgtg gcggctgcgc ccgggccatc
     2821 ctggagaact atatctcagc cctcaacacg ctgtggcatc ctgagtgctt tgtgtgccgg
     2881 gaatgcttca cgccattcgt gaacggcagc ttcttcgagc acgacgggca gccctactgt
     2941 gaggtgcact accacgagcg gcgcggctcg ctgtgttctg gctgccagaa gcccatcacc
     3001 ggccgctgca tcaccgccat ggccaagaag ttccaccccg agcacttcgt ctgtgccttc
     3061 tgcctcaagc agctcaacaa gggcaccttc aaggagcaga acgacaagcc ttactgtcag
     3121 aactgcttcc tcaagctctt ctgctataat cgagctcaag cttcgaattc tgcagtcgac
     3181 ggtaccgcgg gcccgggatc caccggatct agataactga tcataatcag ccataccaca
     3241 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3301 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3361 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3421 ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa
     3481 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3541 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3601 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3661 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3721 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3781 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3841 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3901 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4081 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4141 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4261 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4321 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4381 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4441 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4501 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4561 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4621 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4681 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4741 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4801 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4861 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4921 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4981 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5041 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5101 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5161 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5221 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5281 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5341 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5401 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5461 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5521 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5581 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5641 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5701 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5761 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5821 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5881 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5941 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6001 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6061 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6121 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6181 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6241 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6301 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6361 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6421 aaa
//
