LOCUS       sequence-471088-        6920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Paracingulin (human) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6920)
  AUTHORS   Rouaud F, Huang W, Flinois A, Jain K, Vasileva E, Di Mattia T, 
            Mauperin M, Parry DAD, Dugina V, Chaponnier C, Mean I, Montessuit S,
            Mutero-Maeda A, Yan J, Citi S
  TITLE     Cingulin and paracingulin tether myosins-2 to junctions to 
            mechanoregulate the plasma membrane.
  JOURNAL   J Cell Biol. 2023 Jul 3;222(7):e202208065. doi: 
            10.1083/jcb.202208065. Epub 2023 May 19.
  PUBMED    37204781
REFERENCE   2  (bases 1 to 6920)
  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: "J Cell 
            Biol."; date: "2023-07-3"; volume: "222(7):e202208065. doi"; pages: 
            " 10.1083/jcb.202208065. Epub 2023 May 19"
COMMENT     Sequence Label: Strep-His-Tev-hCGNL1
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  724..947
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1014..1547)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1736..1741,1742..1758,1759..1764))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(1835..1868)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1945..2036
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1961..1978
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(2078..2083,2084..2107,2108..2143,2144..2167)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             2180..2209
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2225..2245
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    6311..6445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6348..6367)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6402..6421
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6566..6731
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattgacttg ggtcgctctt cctgtggatg cgcagatgcc ctgcgtaagc
      721 gggtgtgggc ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa
      781 agtcttaaac tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag
      841 catactggac ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc
      901 gtcgtattaa agaggggcgt ggccaagggc atgtaaagac tatattcgcg gcgttgtgac
      961 aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg
     1021 gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag
     1081 agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc
     1141 gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc
     1201 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
     1261 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
     1321 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
     1381 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
     1441 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
     1501 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
     1561 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     1621 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
     1681 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
     1741 acgaaccgaa caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg
     1801 tcacccggca accttgggca gcagcgaagt cgccataact tcgtatagca tacattatac
     1861 gaagttatct gtaactataa cggtcctaag gtagcgagtt taaacactag tatcgattcg
     1921 cgacctactc cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa
     1981 ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc
     2041 cggattattc ataccgtccc accatcgggc gcggatgtct gcttggtcac accctcaatt
     2101 cgagaaaggc ggcggtagtg gcggcggatc tggcggtagt gcctggagcc atccgcagtt
     2161 tgaaaaggcc ggccatatgc accaccatca tcatcaccat caccatcacg gatctggcgg
     2221 tagtgaaaac ctgtattttc agggcgccgg taccatggaa ctgtacttcg gcgagtacca
     2281 gcacgtgcag caagagtacg gagtccatct ccgtttggca tctgacgata ctcagaaatc
     2341 aaggtccagc caaaactcaa aggccggcag ttacggagtc tcgatccgtg ttcagggtat
     2401 tgacggccac ccatacatcg tgctgaacaa cactgagagg tgcctcgctg gaacaagctt
     2461 ctctgaaaac ggtcctccct tcccaccgcc tgtcattaac aacctgccgc tccattcttc
     2521 aaacggcagt gttcctaagg aaaactcgga ggaattgcag ctgcccgaga acccatacgc
     2581 ccaaccgtcc cctatccgta acctgaaaca gcctctgctc cacgagggaa agaacggtgt
     2641 cctcgataga aaagacggat ccgttaagcc gagccacttg ctgaacttcc agcgccatcc
     2701 tgagctcttg caaccgtacg accctgaaaa gaacgagctc aacttgcaaa accatcagcc
     2761 ctcagaaagt aactggctga agaccctcac tgaggaagga atcaacaaca agaaaccatg
     2821 gacttgcttc cccaagccat cgaactccca gcccacaagc ccatctttgg aggatcctgc
     2881 aaagtccggt gtcactgcga tccgtctctg tagttcggtg gtcattgagg accccaagaa
     2941 acagacgagc gtctgcgtta acgtgcaagg ctgtaccaag gaaagagtgg gagaggaagc
     3001 cttgttcact tctggaaggc ctctgacagc acacagcccg cacgcgcatc ctgagacgaa
     3061 gaaaaccagg cctgacgttc tcccaatcag gaggcaggat agtgctggtc ctgtgttgga
     3121 cggagctagg tctaggagat ccagctcttc aagtaccact ccgacccctg ctaactccct
     3181 gtacaggttc ctgctcgacg atcaggagtg tgccatccac gcagacaacg tgaaccgtca
     3241 tgaaaaccgc cgttacattc cgttcctgcc tggaacgggt agggatatcg acaccggcag
     3301 cattcccgga gtcgatcagc tcatcgagaa attcgaccag aagccaggtt tgcaaaggag
     3361 aggccgttct ggaaagcgca accgtattaa cccggacgat aggaagagat cacgcagtgt
     3421 cgactcagct ttccctttcg gtctccaggg caactccgaa tacttgatcg agttcagcag
     3481 gaacctgggc aagtcgtccg agcacttgct gcgtccttct caggtgtgcc cacaaaggcc
     3541 actgtctcag gaacgccgtg gtaaacaatc agtcggcaga accttcgcta agctccaagg
     3601 tgctgcacac ggagcttctt gtgctcattc acgccctcct cagccaaaca tcgacggcaa
     3661 ggtcctggag acggaaggca gccaggagtc taccgttatt cgtgctcctt ctctcggtgc
     3721 ccaatcaaag aaagaggaag aggttaagac agccacggca accctgatgc tccagaacag
     3781 ggcagcggct acgtcaccag atagtggagc taagaaaatc tctgtgaaga ccttcccctc
     3841 ggcatccaac actcaggcga caccagacct cttgaagggt cagcaagagc tgacgcagca
     3901 aaccaacgaa gagaccgcca aacaaatctt gtacaactac ctgaaggagg gctcaactga
     3961 caacgacgat gctacaaaaa gaaaggtgaa cttggtcttc gaaaaaattc agactctgaa
     4021 gagtcgcgcc gcaggatcgg ctcagggtaa caaccaagcc tgcaacagca cttctgaagt
     4081 gaaggacctg ctcgagcaaa aatctaagct cacaatcgaa gtggctgagt tgcagagaca
     4141 attgcagctg gaggtcaaaa accagcaaaa cattaaggaa gagcgtgaaa ggatgagagc
     4201 caacttggaa gagctgcgct cacagcacaa cgagaaggtc gaagagaact ccacgctcca
     4261 gcaacgtttg gaagagagcg agggagaact gaggaaaaac ctcgaagagt tgttccaagt
     4321 gaagatggag cgtgaacaac accagacaga gatcagggat ctgcaagacc agctctctga
     4381 aatgcatgat gagctggaca gtgctaaaag atcggaagat cgcgagaagg gtgcactgat
     4441 cgaagagttg ctgcaagcga agcaggatct ccaagacctc ttgattgcta aggaaggccg
     4501 tgaggacctg ctccgcaagc gtgagaggga actgacagca ctcaaaggag cgctgaagga
     4561 agaggtcagc tctcacgatc aggagatgga caaactcaag gaacagtacg atgctgagtt
     4621 gcaagccctg cgcgagtccg ttgaagaggc taccaagaac gtcgaagttc tggcctcacg
     4681 tagtaacact agcgaacaag accaggccgg tacagagatg agggttaaat tgctgcagga
     4741 agagaacgag aagctgcaag gcagatccga agagctcgaa aggagagtgg ctcaattgca
     4801 gcgccaaatc gaggatctga agggtgacga agcgaaagct aaggagactc tgaagaaata
     4861 cgagggcgaa attagacagt tggaagaggc actggtgcac gcgcgcaaag aagagaagga
     4921 agccgtctct gcacgccgtg cgctcgagaa cgaattggag ggagctcagg gtaacctgtc
     4981 acaaacaacg caggaacaaa agcagctgtc tgagaaactc aaggaagagt cagaacagaa
     5041 agagcaactc aggagattga agaacgaaat ggagaacgaa agatggcatc tcggcaagac
     5101 catcgaaaaa ttgcagaagg agatggctga tattgtcgaa gcctcgcgca cttccacact
     5161 cgagttgcaa aaccagttgg acgaatacaa agagaagaac cgccgtgaac tggccgagat
     5221 gcaacgtcag ctcaaagaga agactttgga ggctgaaaaa tcccgtctga cagccatgaa
     5281 gatgcaggac ggaatgaggc tgatggaaga ggaactccgt gattaccaga gggctcaaga
     5341 cgaagccctg acgaagcgcc agctcttgga gcaaaccctg aaggacctcg agtacgaact
     5401 cgaggctaaa agtcacttga aggacgatag atcgcgcctg gttaaacaga tggaggctaa
     5461 ggtgtcccaa ttggaaatgg agctggagga agagagaaac aactccgacc tgctcagcga
     5521 gcgcatctcg cgttccaggg aacagatgga gcaactgaga aacggattgc tgcaggaacg
     5581 tgcggctagg caagatctgg aatgcggcaa gattagcctc gagagacaga acaaagactt
     5641 gaagagtcgc atcattcacc tggagggatc gtacaggtct agtaaggaag gtctggttgt
     5701 gcaaatggag gctcgtatcg ccgaactcga ggataggttg gaaagtgaag agagagaccg
     5761 cgctaacctg cagctctcca acaggagact cgaacgtaaa gtcaaggagt tggttatgca
     5821 agtggacgat gagcacttgt ctctgactga tcagaaggac caactctcat tgagactgaa
     5881 agcaatgaag cgccaggtgg aagaggcgga agaggaaatc gaccgtctgg aatcgtccaa
     5941 gaaaaagctg cagagggagc tcgaggaaca aatggatatg aacgaacatt tgcagggtca
     6001 gctgaactca atgaaaaagg acctccgctt gaaaaagttg cccagtaagg tgctggacga
     6061 tatggacgat gacgatgacc tgtccacaga tggtggcagc ctctacgagg ctccagttag
     6121 ctacacgttc agcaaggact ctaccgtggc ctctcagatc tctagagcct gcagtctcga
     6181 caagcttgtc gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt
     6241 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     6301 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6361 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6421 caatgtatct tatcatgtct ggatctgatc actgcttgag cctagaagat ccggctgcta
     6481 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctatcataac
     6541 ccctagggta tacccatcta attggaacca gataagtgaa atctagttcc aaactatttt
     6601 gtcattttta attttcgtat tagcttacga cgctacaccc agttcccatc tattttgtca
     6661 ctcttcccta aataatcctt aaaaactcca tttccacccc tcccagttcc caactatttt
     6721 gtccgcccac aaccggttga cttgggtcaa ctgtcagacc aagtttactc atatatactt
     6781 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6841 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6901 gaaaagatca aaggatcttc
//
