LOCUS       sequence-365837-        5939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expressing recombinant p130Cas in insect cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5939)
  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 5939)
  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: p130Cas Y15F
FEATURES             Location/Qualifiers
     source          1..5939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     375..394
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     628..645
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  779..1003
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1070..1603)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1773..1778,1779..1795,1796..1801))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2153..2244
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2169..2186
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2311..2328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2350..2370
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3697..3831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3734..3753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3788..3807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3860..4025
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4209..4663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4296..4315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4505..4526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4689..4793
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4794..4862,4863..5654)
                     /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     complement(5012..5031)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5825..5939,1..474)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
       61 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
      121 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
      181 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
      241 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
      301 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
      361 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
      421 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
      481 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
      541 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      601 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      661 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agaccagccg
      721 cgtaacctgg caaaatcggt tacggttgag taataaatgg atgccctgcg taagcgggtg
      781 tgggcggaca ataaagtctt aaactgaaca aaatagatct aaactatgac aataaagtct
      841 taaactagac agaatagttg taaactgaaa tcagtccagt tatgctgtga aaaagcatac
      901 tggacttttg ttatggctaa agcaaactct tcattttctg aagtgcaaat tgcccgtcgt
      961 attaaagagg ggcgtggcca agggcatggt aaagactata ttcgcggcgt tgtgacaatt
     1021 taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg
     1081 tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag
     1141 ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt
     1201 tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc
     1261 cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa
     1321 cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg
     1381 tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg
     1441 ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt
     1501 cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag
     1561 ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg
     1621 cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac
     1681 aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc
     1741 agttgcgtga gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca
     1801 actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca
     1861 gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc
     1921 atcgtcaggc attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctgc
     1981 cctggcttca ggagatcgga agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc
     2041 cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg
     2101 actctagcta tagttctagt ggttggctac gtatactccg gaatattaat agatcatgga
     2161 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
     2221 tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc
     2281 ggatctcggt ccgaaaccat gtcgtactac catcaccatc accatcacga ttacgatatc
     2341 ccaacgaccg aaaacctgta ttttcagggc gccatggatc cggaattcgc aaacagcctg
     2401 gagcctgaac cttggttctt caagaatctg agccgtaagg acgccgagcg gcagcttttg
     2461 gcgcccggga acacgcatgg atccttcctg atccgggaaa gcgaaagcac tgcggggtcc
     2521 ttttccctgt cggtcagaga cttcgaccag aaccagggag aagtggtgaa acattacaag
     2581 atccgtaacc tagacaacgg tggcttctac atctcccctc gtatcacttt tcccggattg
     2641 cacgatctag tccgccatta caccaacgcc tctgatgggc tgtgcacaaa gttgagccgt
     2701 ccttgccaga cccagaagcc ccagaaacca tggtgggagg acgaatggga agttcccagg
     2761 gaaacactga agttggtgga gcggctggga gctggccagt tcggggaagt gtggatgggg
     2821 tactacaacg gacacacgaa ggtggcggtg aagagtctga aacaagggag catgtccccc
     2881 gacgccttcc tggctgaggc taacctcatg aagcagctgc agcacccgcg gctagtccgg
     2941 ctttatgcag tggtcaccca ggaacccatc tacatcatca cggaatacat ggagaacggg
     3001 agcctagtag attttctcaa gactccctcg ggcatcaagt tgaatgtcaa caaacttttg
     3061 gacatggcag cccagattgc agagggcatg gcgttcatcg aagaacagaa ttacatccat
     3121 cgggacctgc gcgccgccaa catcctggtg tctgacacgc tgagctgcaa gattgcagac
     3181 tttggcctgg cgcgcctcat tgaggacaat gagtacacgg cccgggaggg ggccaaattt
     3241 cccattaagt ggacagcacc agaagccatt aactatggga ccttcaccat caagtcagac
     3301 gtgtggtcct tcgggatctt gcttacagag atcgtcaccc acggtcgaat cccttaccca
     3361 ggaatgacca accctgaagt cattcagaac ctggagagag gctaccgcat ggtgagacct
     3421 gacaactgtc cggaagagct gtaccacctc atgatgctgt gctggaagga gcgcccagag
     3481 gaccggccca cgtttgacta ccttcggagt gttctggatg acttcttcac agccacagag
     3541 ggccagttcc agccccagcc ttgatagaag cttgtcgaga agtactagag gatcataatc
     3601 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3661 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3721 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3781 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg
     3841 cttgagccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
     3901 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     3961 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     4021 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     4081 tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat
     4141 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg
     4201 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4261 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4501 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4561 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4621 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     4681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     4801 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4861 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4921 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4981 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5041 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5101 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5161 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5221 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5281 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5341 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5401 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5461 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5521 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5581 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5641 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5701 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5761 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5821 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5881 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaact
//
