LOCUS       sequence-471086-        6623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Cingulin (human) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6623)
  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 6623)
  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-hCGN
FEATURES             Location/Qualifiers
     source          1..6623
                     /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"
     RBS             3637..3645
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    6014..6148
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6051..6070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6105..6124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6269..6434
                     /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 caggcaccta acatggctga
     2281 gccaagagga cctgtggatc acggagtcca aatccgcttc attaccgaac ctgtgtctgg
     2341 agctgagatg ggtactctgc gccgtggtgg caggagaccc gctaaggacg cccgtgcatc
     2401 aacttacgga gttgctgtga gggtccaagg aatcgccggt cagcccttcg tggtcctgaa
     2461 ctcaggagag aagggaggtg acagtttcgg tgtgcagatt aaaggcgcca acgatcaagg
     2521 tgcatctgga gcactgtcct ctgacctcga attgcctgag aacccctact ctcaggtcaa
     2581 gggtttccct gccccctcgc aatcttcaac ctccgacgag gaaccaggag cttactggaa
     2641 cggcaagctg ctccgttctc actctcaagc ttctctggct ggaccaggac ctgtggatcc
     2701 gtccaacagg agtaactcga tgctggagct cgctccaaag gtcgcctcac cgggcagtac
     2761 aatcgacacg gctcctttga gttcggttga ttcgctgatt aacaagttcg actcccaact
     2821 gggcggacaa gcacgtggta ggactggtag gaggactagg atgctccctc ccgagcagcg
     2881 taagaggtcc aaaagcttgg attcgagact gccacgcgac acattcgagg aaagagaacg
     2941 ccagtccacg aaccactgga cctccagcac taagtacgac aaccatgtgg gcacctcgaa
     3001 acagccagct cagtcccaaa acctgtcgcc gctctccgga ttctctcgtt caaggcagac
     3061 tcaagattgg gtcttgcaga gcttcgagga acctaggaga tctgctcaag accccaccat
     3121 gctgcagttc aagtcgactc ctgacttgct gcgtgatcag caagaggctg ccccaccggg
     3181 ttctgttgac cacatgaagg ccaccatcta cggcattctg agggagggat cttcagagag
     3241 cgaaacttct gtccgccgta aggtgtcatt ggtcctggaa aaaatgcagc ccctcgttat
     3301 ggtgagttcg ggcagtacaa aggcggttgc tggtcagggc gagttgacgc gtaaggtgga
     3361 ggaactccaa aggaaattgg acgaggaagt caagaaacgc caaaaactgg aaccatcaca
     3421 ggttggcctg gagagacaac tcgaggaaaa gactgaggaa tgctcccgcc tgcaggaact
     3481 cttggagagg agaaagggag aagctcagca atctaacaag gagctccaga acatgaaaag
     3541 actgctcgac caaggagagg atttgcgcca cggtctggaa acacaagtga tggagctcca
     3601 gaacaagttg aaacatgtgc agggtcctga acccgctaag gaggtcttgc tgaaagacct
     3661 cttggaaaca agagagctgc tcgaggaagt gctggagggc aagcaacgcg tcgaggaaca
     3721 gctcagattg cgcgagcgtg aattgacggc actgaaggga gcgctgaaag aggaagtcgc
     3781 ttcccgtgat caggaggttg aacacgtgcg ccagcaatac caacgtgaca ccgagcagct
     3841 ccgccgttcc atgcaagacg ctactcagga tcatgctgtg ttggaggccg aacgccagaa
     3901 gatgagcgcc ctggtcagag gactccaacg cgagttggag gaaacctcag aggaaactgg
     3961 tcactggcaa agtatgttcc agaagaacaa agaagatctg cgtgccacca agcaagaatt
     4021 gctgcagctc aggatggaga aagaggaaat ggaggaagag ctgggtgaaa agatcgaggt
     4081 gctgcaaaga gaactggagc aagcaagggc atctgctggc gacacacgtc aggtcgaagt
     4141 tctgaagaaa gagctcttga ggacgcagga agagctcaag gaattgcaag cagagagaca
     4201 gtcacaagag gtcgcgggta ggcatagaga tcgcgagctg gaaaagcagc tggctgtcct
     4261 ccgcgttgaa gccgaccgtg gcagggagct cgaagagcag aacctgcagc tccaaaagac
     4321 tttgcagcaa ctgcgtcaag attgtgaaga ggcctccaag gcaaaaatgg ttgctgaggc
     4381 cgaagcaaca gtgttgggtc agaggagagc agcggtggaa accactctgc gcgagacgca
     4441 agaagagaac gacgagttcc gccgtaggat cttgggtctg gaacagcaac tcaaggagac
     4501 ccgtggcttg gttgatggtg gcgaagcggt ggaggctaga ctgcgcgaca agctccagag
     4561 gttggaggct gaaaaacagc aattggaaga ggccctgaac gcaagccaag aagaggaagg
     4621 ttctctggct gccgcaaaga gggcgctcga ggctagattg gaggaagcac agcgtggtct
     4681 ggcgaggctc ggccaagaac agcaaacact gaaccgtgct ctcgaggaag agggcaagca
     4741 gcgtgaagtg ctgagacgcg gaaaggctga gctcgaagag caaaaaagac tgctcgaccg
     4801 caccgtcgat cgtttgaaca aggagctgga aaaaatcggc gaggatagca agcaggcatt
     4861 gcagcaactg caggcgcaac tggaagacta caaggagaaa gctcgtaggg aggtggcgga
     4921 tgctcagaga caagccaagg actgggcctc cgaggcagaa aaaacctcag gaggtctcag
     4981 tcgcttgcaa gacgagattc agcgtctcag gcaggcgttg caagctagcc aggccgagag
     5041 ggatactgct agactggaca aggaattgct ggcccaaaga ctgcagggac tcgagcagga
     5101 agcagagaac aagaaacgct ctcaggacga tagagcccgc caattgaagg gtctggaaga
     5161 gaaagtctca cgtctggaaa cagagctcga tgaagagaag aacacagtcg agctcttgac
     5221 ggaccgtgtt aaccgtggaa gggaccaagt ggatcagttg aggaccgaac tgatgcagga
     5281 gcgtagcgcc aggcaagact tggaatgcga taagatctct ctggagagac agaacaagga
     5341 cctcaaaact cgcttggcat ccagcgaggg tttccagaag cccagtgcgt cgctctccca
     5401 attggaaagc cagaaccaac tgctccaaga gcgtctgcag gctgaagaga gggaaaagac
     5461 cgttctccag tcaactaacc gcaaattgga gcgtaaggtg aaagagctga gtatccagat
     5521 tgaagacgag cgtcaacacg tgaacgacca aaaggatcag ctctctttga gagtcaaggc
     5581 actgaaacgc caggttgatg aagcggaaga ggaaatcgag cgcctggacg gcctccgtaa
     5641 gaaagcccag agggaagtcg aggaacaaca tgaagttaac gagcagctcc aagctcgcat
     5701 taagagcttg gaaaaagact cttggaggaa agctagcaga tctgcggctg aatcagccct
     5761 gaagaacgag ggactctctt cagacgagga attcgatagt gtctacgacc caagttcgat
     5821 cgcttcattg ctgacagaga gtaacctgca gacgtccagc tgttctagag cctgcagtct
     5881 cgacaagctt gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt
     5941 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     6001 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6121 catcaatgta tcttatcatg tctggatctg atcactgctt gagcctagaa gatccggctg
     6181 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactatcat
     6241 aacccctagg gtatacccat ctaattggaa ccagataagt gaaatctagt tccaaactat
     6301 tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc atctattttg
     6361 tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt tcccaactat
     6421 tttgtccgcc cacaaccggt tgacttgggt caactgtcag accaagttta ctcatatata
     6481 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6541 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6601 gtagaaaaga tcaaaggatc ttc
//
