LOCUS       sequence-365815-        7464 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of recombinant p130Cas in SF9 cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7464)
  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 7464)
  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
FEATURES             Location/Qualifiers
     source          1..7464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(92..97,98..114,115..120))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        472..563
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     488..505
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             630..647
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             669..689
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3522..3656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3559..3578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3613..3632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3685..3850
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4034..4488
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4121..4140)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4330..4351
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4514..4618
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4619..4687,4688..5479)
                     /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(4837..4856)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5650..6238
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6139..6158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6392..6409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6543..6767
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6834..7367)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
       61 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
      121 ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
      181 cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca
      241 tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc
      301 ctggcttcag gagatcggaa gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
      361 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
      421 ctctagctat agttctagtg gttggctacg tatactccgg aatattaata gatcatggag
      481 ataattaaaa tgataaccat ctcgcaaata aataagtatt ttactgtttt cgtaacagtt
      541 ttgtaataaa aaaacctata aatattccgg attattcata ccgtcccacc atcgggcgcg
      601 gatctcggtc cgaaaccatg tcgtactacc atcaccatca ccatcacgat tacgatatcc
      661 caacgaccga aaacctgtat tttcagggcg ccatgggatc cggcaccatg aagtacctga
      721 acgtgctggc caaagccctc tatgacaacg tagctgagtc tccggatgaa ctctccttcc
      781 gaaagggtga catcatgaca gtgctcgagc gggacactca gggcctggat ggctggtggc
      841 tctgctcact gcatgggcgc cagggcattg tgcctggtaa ccgcctcaag attctggttg
      901 gcatgtatga caagaagcca gcagcacctg gaccaggccc acctgccacc ccaccccagc
      961 cccagcccag ccttccccag ggcgtccata ctccagtgcc tccagcttcc cagtacagtc
     1021 ccatgctccc cactgcatac caaccccaac ctgacaatgt gtacctggta cccactccca
     1081 gcaagactca gcaaggtctc taccaagccc ctgggcccaa cccacagttc cagtcacccc
     1141 cagccaaaca aacatccaca ttctcaaagc aaacacctca tcactcattt cccagccctg
     1201 ccacagacct ttaccaggtg cctccagggc ctggaagtcc agcccaggac atttaccagg
     1261 tgccaccttc cgctggcaca gggcatgaca tctaccaagt ccctccatct ttggacaccc
     1321 gcagctggga gggaacgaag ccaccggcaa aggtggtggt acctactcga gtgggacagg
     1381 gctatgtgta cgaggcctcc caggcagaac aggatgagta cgacaccccg cgccacctgc
     1441 tagctccagg ttcccaggac atctatgatg tgccccctgt tcgaggactg cttcccaacc
     1501 agtatggcca ggaggtatat gacaccccgc ctatggcagt caaaggcccc aatggccgag
     1561 acccgttgtt ggatgtgtat gacgtgcccc ctagtgtgga gaaaggcctg ccgccgtcca
     1621 accatcattc ggtgtatgat gttcctcctt ctgtgagcaa ggatgtgcct gatggcccac
     1681 tgctgcgtga ggaaacatat gatgtacccc ctgccttcgc caaacccaag ccttttgatc
     1741 cgacccgaca tccattaatc ctcgctgcac ctcctccgga ctccccacca gctgaggatg
     1801 tgtatgatgt gccaccccct gctcctgacc tctatgatgt gccccctggt ttgcggcggc
     1861 ctggccctgg aacactgtat gacgtgcctc gagagcgggt gctccctcct gaggtggctg
     1921 atgggagtgt aattgatgat ggtgtgtatg ctgtgccccc accagccgag cgagaggcgc
     1981 caacagatgg caagcgcctg tcggcctcta gcacgggcag cacacgcagc agccaatcgg
     2041 cgtcttcctt ggaggtagtg gtgccaggtc gggagcccct ggagctggaa gttgctgtgg
     2101 agaccctggc tcggctacag cagggtgtga gcaccaccgt agcccacctt ctggacctgg
     2161 tgggcagtgc cagtggcccc gggggctggc gtagtacctc tgagccccag gagcctcccg
     2221 tgcaggacct gaaagctgca gtggccgcag ttcatggggc agtacatgag ctcctggagt
     2281 ttgcccgcag tgcagtgagc agtgccactc acacttctga ccgcactttg cacgccaagc
     2341 ttagccggca gctacagaag atggaggatg tgtaccagac actggtggtc catggtcagg
     2401 tccttgacag tggccgggga ggtccaggat tcactctgga cgacctggac cgcctggtgg
     2461 cctgctcacg ggctgtgccc gaggatgcca agcagctggc ctcctttttg catggtaatg
     2521 cctcgctgct tttcagacgg accaaagccc ctggccccgg gcctgagggc agcagctccc
     2581 tgcacctcaa ccccaccgat aaagccagca gcatccagtc acgccctcta ccctcacctc
     2641 caaagttcac ctcccaggac tctccggatg gccagtatga gaacagtgaa gggggttgga
     2701 tggaggacta tgactacgtt catctgcagg ggaaggagga atttgagaag acccagaagg
     2761 aactgctgga aaagggcaac atcgtgcgac agggaaaagg ccaactggag ctgcagcagc
     2821 tgaaacagtt tgagcgactg gagcaggagg tgtctcgtcc catagaccat gacctggcca
     2881 actggacacc agcccagccc cttgtgccgg ggcggacagg gggactgggg ccttcagacc
     2941 gacagctgct gctcttctac ttggagcagt gtgaggccaa cctgaccaca ctgacagatg
     3001 cggtggacgc cttcttcacc gcagtggcca ccaaccagcc acccaagatc tttgtggcac
     3061 acagcaagtt tgtcattctt agtgcacaca agcttgtgtt cattggggac acactgtcac
     3121 ggcaggcaaa ggcagctgat gtgcgaagcc aagtgaccca ctacagcaat ctgctgtgtg
     3181 acctcctgcg tggcatcgtg gccaccacca aggccgctgc cctgcagtac ccatccccat
     3241 ctgctgccca ggacatggta gacagggtca aggagctagg ccacagcact cagcagttcc
     3301 gccgtgtcct gggccagctg gctgccgcct gaaagcagaa gactgcggcc gctttcgaat
     3361 ctagagcctg cagtctcgag gcatgcggta ccaagcttgt cgagaagtac tagaggatca
     3421 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3481 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3541 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3601 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     3661 cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg
     3721 tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac
     3781 tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc aactattttg
     3841 tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact
     3901 ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct
     3961 gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg
     4021 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4081 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4141 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4201 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4261 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4321 ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4381 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4441 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     4501 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4561 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4621 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4681 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4741 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4801 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4861 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4921 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4981 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5041 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5101 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5161 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5221 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5281 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5341 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5401 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5461 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5521 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5581 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5641 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5701 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5761 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     5821 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5881 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5941 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6001 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6061 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6241 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6301 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6361 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6421 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca
     6481 gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg
     6541 ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa
     6601 gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc
     6661 atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg
     6721 tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac
     6781 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     6841 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     6901 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     6961 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
     7021 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     7081 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     7141 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     7201 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     7261 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     7321 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
     7381 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     7441 ctgtacaaaa aaacagtcat aaca
//
