LOCUS       sequence-469647-        9042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Cingulin (mouse) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9042)
  AUTHORS   Vasileva E, Spadaro D, Rouaud F, King JM, Flinois A, Shah J, 
            Sluysmans S, Mean I, Jond L, Turner JR, Citi S
  TITLE     Cingulin binds to the ZU5 domain of scaffolding protein ZO-1 to 
            promote its extended conformation, stabilization, and tight junction
            accumulation.
  JOURNAL   J Biol Chem. 2022 Apr;298(4):101797. doi: 10.1016/j.jbc.2022.101797.
            Epub 2022 Mar 5.
  PUBMED    35259394
REFERENCE   2  (bases 1 to 9042)
  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 Biol 
            Chem."; date: "2022-04"; volume: "298(4):101797. doi"; pages: " 
            10.1016/j.jbc.2022.101797. Epub 2022 Mar 5"
COMMENT     Sequence Label: mCGN-Myc-His
FEATURES             Location/Qualifiers
     source          1..9042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(111..130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     321..342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(461..481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        466..795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      646..781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     708..727
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             862..1656
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(916..935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1526..1545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1830..1951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1867..1886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1921..1940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2000..2016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2000..2016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2013..2035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2024..2040
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2048..2054,2055..2072,2073..2078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2093..2114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2231..2248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2402..2990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2482..2501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3161..3952,3953..4021))
                     /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     3784..3803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4022..4126)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4201..4220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4392..4771
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4772..4975
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4926..4946
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5020..5039
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5020..5038
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             8684..8713
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             8729..8746
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(8790..8807)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8796..9020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
       61 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
      361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
      421 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
      481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
      541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
      601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
      661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
      721 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
      781 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
      841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
      901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
      961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     1021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     1081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     1141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     1201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     1261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     1321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     1381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     1441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     1501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     1561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     1621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     1681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     1741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     1801 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     1861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     1981 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4141 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     4201 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     4261 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     4321 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     4381 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     4441 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     4501 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     4561 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     4621 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     4681 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     4741 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     4801 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     4861 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     4921 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     4981 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     5041 gacccaagct ggctagcgtt taaacgggcc ctctagactc gaggccacca tgatggctga
     5101 tccccggggc cccgtagacc atggagttca aattcgattt attacagagc cagagggtgc
     5161 tactgaaatg ggcacccttc gtcgtagtgg gcggcgccct gcaagggatg caagagccag
     5221 tacctatggg gtcgctgtgc gtgtgcaggg catcgccggg caaccgtttg tggttcttaa
     5281 cagtggagag aaagggactg actcctttgg ggtccagatc aaagggggca acaaccgggg
     5341 gtccccaggt gcactgagct ccgattcaga gctccccgaa aacccctact ctcaagtcaa
     5401 gggacgtcct gccacttctc ggagcagcac atctgatgag gagcccaagg accacttgaa
     5461 tggaaaactg atccgttccc agtcgcaagc ctctctgacc gggcttgcct ttatgagtcc
     5521 tagtaatagg agcaccagct tgctggaact agcccctaaa ccgacttcct cgattaacac
     5581 cattgacact gcccccttgt cttcagtgga ctcgctcatc aacaagtttg atagtcagaa
     5641 gggggggcag gtccggggtc gcactgggcg ccgcacaagg acacttcccc acgaacagcg
     5701 caagcggagc cagagtctgg acagcaggct cccacgggat acacgtgagg aacgggagca
     5761 ccagtccgct aaccactgga cccgcggcac aaagtatgac aaccatgtgg acagttcaaa
     5821 gaacccatcg cagaagcaga gcccgttcag cagcttcagc cgttcgcgtc agactcaaga
     5881 ctgggtgctt cagagttttg aggagacacg ggatccagcc atggtgcagt tcaaatcaac
     5941 tccagacctt ctcagagacc agagagagac agcgccacca ggcagtgcgg accatgtgaa
     6001 ggccaccatc tatggcatcc taagggaggg aagctcggag agtgaggcat ctgtgaggag
     6061 gaaagttagt ttggtgctgg agcaaatgca gcctctgggg atggtttctc ctgcttctac
     6121 caaggccctg gcaggacagg ctgagcttac ccgaaaaatg gaggaacttc agaagaagct
     6181 ggatgaggag gtgaagaagc ggcagaagct agaaccatcc agggttgggc tggagaggca
     6241 gctggaggag aaggctgaag agtgccaccg actgcaggag ttgctggaga ggagaaaggg
     6301 ggaggtccag cagagcagca aggaacttca gaacatgaag ctcctgctag gccaggaaga
     6361 aggactacgg catgggctgg aggcccaggt gaaggagcta cagcttaagc tcaagcatag
     6421 ccagagtccc gattctggaa aggaaagtct cctaaaggac ttgttagaca cccgggaact
     6481 tctagaagaa ctcttagagg gcaaacaacg agtagaggag cagctcaggc tgagggaacg
     6541 ggagctgaca gccctgaagg gggccctgaa agaggaggtg gcctcccacg accaggaggt
     6601 ggaacatgtc cggctgcagt accagcgaga cacggagcag cttcgccgga gcatgcaaga
     6661 tgcgacccag gaccatgctg cactggaggc tgagaggcag aagatgtctt ccctcgtgcg
     6721 ggagctgcaa agagagctag aggagacctc tgaggagaca gggcattggc agagcatgtt
     6781 ccagaagaac aaggaagagc tgagagccac caagcaggaa ctgctgcaac tgcggatgga
     6841 gaaggaggag atggaagagg aacttgggga gaagatggag gtcttacaga gggacttaga
     6901 gcaggcccga gccagtacaa gagatactca ccaggttgag gaacttaaaa aggagcttcg
     6961 acgcacccag ggggagctta aagagcttca ggcagagcag cagaaccagg aggtgactgg
     7021 gcgacaccag aaccaggtgt tagagaagca gctggcagcg ctgagggagg aggctgatcg
     7081 aggccgagag ctggagcagc agaacctcca gctacaaaag accctgcagc aactgcggca
     7141 ggactgtgaa gaggcttcca aggctaaggt ggcatcagag acagaggcca tggtgctggg
     7201 gcagcgtcgg gcaaccgtgg aaaccacact tcgggagaca caagaggaaa atgacgaatt
     7261 ccgaaggcgc atcctgggtc tggagcagca gttgaaggag gcccgaggtc tggcggaagg
     7321 tggggaagcg gtggaggcac ggcttcggga caaagtgcat cggctggagg tagagaaaca
     7381 gcagctggaa gaggctttga atgctgcccg ggaggaggag gggaacctgg cagcagccaa
     7441 gcgggctctg gaggtccggc tagatgaggc ccagcggggg ctggcacgcc ttgggcagga
     7501 acagcaagca ctgaaccggg ccctagagga agaggggaag cagcgggagg ctctccggag
     7561 gagcaaggca gagttggagg agcagaagcg tttgctgaac aggactgtag accgactcaa
     7621 caaagagctg gagcagattg gggatgactc caagctagcc ctgcagcagc tccaggctca
     7681 gatggaagat tacaaggaaa aggcccggaa ggaagtagct gatgcccagc gtcaggccaa
     7741 ggattgggcc agtgaggctg aaaagaattc aggggggctg agccggcttc aggatgagct
     7801 gcaaaggctg cggcaagccc tgcagacatc ccaggctgag cgggacacag ccaggctgga
     7861 caaagagctg ctggcccagc gactgcaggg tctagagcaa gaggctgaga acaagaaacg
     7921 cttccaggac gacaaggctc gacagctgaa gagcctagag gagaaagtct cgcggctgga
     7981 agcggagtta gatgaagaaa agaacactgt ggaactgtta acagatcggg tgaaccgtgg
     8041 ccgggatcag gtggatcaac tgaggacaga gctcatgcag gagagatctg ctcgacagga
     8101 cttggagtgt gacaaaatct ccttggagag acagaacaag gatttgaaga cccggttggc
     8161 cagttcagag ggcttccaga aacccagcgc tagcctctcc cagcttgagt cccagaatca
     8221 gctgctgcaa gagcggctgc aagctgaaga gagagagaag acagttctgc agtccaccaa
     8281 tcgcaaacta gaacggaggg ttaaggagct ctccatccag attgatgatg aacggcagca
     8341 tgtcaacgac cagaaggacc agctaaccct gagggtgaag gctttgaagc ggcaggtgga
     8401 tgaagcagaa gaggaaattg agcggctgga cagcctgagg aagaaggcac agcgggaact
     8461 ggaggaacag catgaggtca atgaacagct ccaagcccgg atcaaatcct tggaaaagga
     8521 tgcctggcgg aaagcttccc gctcagctgc tgagtcagcc ctcaaacaag aggggctgag
     8581 ctctgatgag gagtttgaca acgtctacga cccctcatcc attgcatcac tgcttactga
     8641 gagcaacctg cagactagct catgtaggta ccaagcttac gtagaacaaa aactcatctc
     8701 agaagaggat ctgaatagcg ccgtcgacca tcatcatcat catcattgag tttaaacggt
     8761 ctccagctta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     8821 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     8881 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     8941 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     9001 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc ag
//
