LOCUS       sequence-365924-        7363 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 7363)
  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 7363)
  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 DN
FEATURES             Location/Qualifiers
     source          1..7363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(609..611,612..614,615..1325)
                     /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(654..675)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(915..934)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1262..1283
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4147..4268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4184..4203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4238..4257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4275..4730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4412..4433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4624..4643
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4757..4861
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4863..5220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4863..5054
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4886..4906)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5071..5206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5255..6049
                     /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             5255..6049
                     /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(5309..5328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5919..5938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6237..6256)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6281..6328
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6657..7245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7146..7165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      601 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      661 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      721 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      781 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      841 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      901 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      961 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1021 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1081 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1141 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1201 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1261 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1321 acaagtccgg actcagatct cgagctatgg acgatcttga cgctctcttg gcagatttgg
     1381 aaagtacgac tagccacata tcaaaacggc cggtcttcct gagcgaggag acaccatatt
     1441 cctatcccac cggaaaccat acgtaccaag aaattgctgt gccgcctcca gttccacctc
     1501 ctccgagttc tgaagcgttg aatggtacca tactcgatcc cctcgaccaa tggcagccat
     1561 caggctctag gtttattcac cagcagccgc agtcatcaag ccctgtctat gggtcctctg
     1621 cgaaaacgag ttcagtttca aacccgcaag atagtgtcgg ttcaccttgc tcacgagtcg
     1681 gtgaggaaga gcatgtgtat tcatttccga ataagcagaa aagtgcggag ccatctccca
     1741 cggttatgtc aacctctctc ggttctaacc tgagcgaact ggatcgcctc cttctggaac
     1801 tgaatgcagt acagcataat ccacccggtt tcccggccga tgaagccaat agtagccctc
     1861 cattgcctgg ggcattgtct cccctgtacg gggtccctga aacgaatagc ccattgggag
     1921 ggaaagcggg gccgctcacg aaagagaaac ccaagcgcaa tggtggacgg ggtctggagg
     1981 acgttcggcc gtccgtagag tccctgttgg atgaactgga atcatcagta ccgagtcctg
     2041 ttccggcaat taccgtgaat caaggcgaga tgtcctctcc ccagcgagtc acgagcacgc
     2101 agcaacaaac gcgcatcagc gcgagcagcg ctacgagaga acttgacgag cttatggcct
     2161 cccttagcga tttcaaattc atggcacaag gaaagactgg ttccagcagc ccgccgggtg
     2221 gtccaccaaa accggggtcc caactcgata gtatgctggg ttcactccag agtgatctga
     2281 ataagctcgg cgtagctacc gtggctaaaa tggacgacct ggacgcactg ctcgcagatc
     2341 tcgaatcaac gacgtcacat atttccaagc gccccgtctt tcttagtgag gagaccccat
     2401 acagctaccc aacaggtaac cacacgtatc aggagattgc ggttcctcct ccagtcccac
     2461 ctccaccgtc cagcgaggcg ctgaacggaa caatactcga ccctctcgac caatggcagc
     2521 ctagtggttc cagatttata catcagcagc ctcaaagcag tagcccggtg tatgggtcta
     2581 gtgctaaaac atcttccgta tccaaccccc aagatagcgt gggttcaccg tgctcccggg
     2641 tgggagaaga agaacatgtt tacagttttc ccaacaagca aaagagtgcg gagccgtcac
     2701 caacagtgat gtctacgtct ctgggctcta acctttctga gctggatagg ctgttgcttg
     2761 aactcaacgc ggtgcagcac aacccgccgg ggttcccagc agatgaagcc aatagcagtc
     2821 ccccactccc aggggctctt agtccattgt atggtgttcc ggagacaaac tcccccctgg
     2881 gaggcaaggc cgggccactg acgaaagaga aaccaaagcg caatggtggc cgaggattgg
     2941 aagatgttag gcctagtgtt gaatctctct tggatgaact ggagagttct gtgccgagtc
     3001 ccgtacctgc catcaccgtt aatcagggag agatgagttc ccctcaacgg gtaacgtcaa
     3061 cgcaacaaca gactcgcata agtgcttcat ccgccacgcg ggaattggac gagctgatgg
     3121 cctcactcag cgacttcaaa tttatggctc aggggaagac cggttcatcc agccctcctg
     3181 gaggtccacc aaaacccggc tctcaacttg atagcatgtt ggggagcctg cagagtgacc
     3241 ttaacaagct tggagtagcg acggtcgcga agggcgtgtg tggtgcttgc aaaaagccta
     3301 tcgctggtca agtagtcacg gcaatgggta agacgtggca cccagagcat tttgtctgca
     3361 cccactgcca agaagagata ggctcaagga atttcttcga aagagacgga cagccctatt
     3421 gcgagaaaga ttaccacaat cttttcagtc caaggtgcta ctactgtaac ggccccattc
     3481 tcgataaggt ggtaaccgcg cttgacagga cgtggcatcc tgaacatttt ttttgcgctc
     3541 aatgcggcgc gttttttggt ccggagggct tccacgagaa agatggaaag gcttattgca
     3601 ggaaggatta tttcgacatg ttcgctccca agtgtggggg gtgtgctcgg gctattctag
     3661 agaattacat cagcgcactt aacacgttgt ggcacccgga gtgctttgtc tgccgcgagt
     3721 gttttactcc ttttgtcaat gggagtttct ttgaacatga cggccagccg tattgtgagg
     3781 ttcattacca tgagcgccga gggtcactgt gcagcgggtg ccagaagcct ataaccggga
     3841 gatgcatcac tgcaatggca aaaaaatttc atccagaaca cttcgtctgc gcgttttgct
     3901 tgaaacaatt gaacaaagga acatttaaag aacaaaatga taagccttac tgtcagaatt
     3961 gcttccttaa attgttctgc tagtgagaat tctgcagtcg acggtaccgc gggcccggga
     4021 tccaccggat ctagataact gatcataatc agccatacca catttgtaga ggttttactt
     4081 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4141 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4261 gtatcttaac gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4321 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4381 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4441 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4501 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4561 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4621 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4681 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4741 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4801 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4861 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4921 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5101 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5161 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5221 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5281 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5341 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5401 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5461 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5521 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5581 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5641 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5701 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5761 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5821 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5881 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5941 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6001 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6061 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6121 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6181 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     6241 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     6301 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6361 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6421 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6481 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7261 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7321 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agt
//
