LOCUS       sequence-469307-        8901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression of Paracingulin (dog) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8901)
  AUTHORS   Flinois A, Mean I, Mutero-Maeda A, Guillemot L, Citi S
  TITLE     Paracingulin recruits CAMSAP3 to tight junctions and regulates 
            microtubule and polarized epithelial cell organization.
  JOURNAL   J Cell Sci. 2024 Mar 1;137(5):jcs260745. doi: 10.1242/jcs.260745. 
            Epub 2023 May 15.
  PUBMED    37013686
REFERENCE   2  (bases 1 to 8901)
  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 
            Sci."; date: "2024-03-1"; volume: "137(5):jcs260745. doi"; pages: " 
            10.1242/jcs.260745. Epub 2023 May 15"
COMMENT     Sequence Label: cCGNL1-HA
FEATURES             Location/Qualifiers
     source          1..8901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(304..315)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3895..3921
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3969..3986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3975..4199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4245..4673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4332..4351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4542..4563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4682..4702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4687..5016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4867..5002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5064..5111
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5130..5504
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5634..5755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5671..5690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5725..5744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5804..5820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5804..5820)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5817..5839)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5828..5844
                     /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(5852..5858,5859..5876,5877..5882))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5897..5918
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6035..6052)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6286..6305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6965..7756,7757..7825))
                     /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     7588..7607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7826..7930)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8005..8024)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8196..8575
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8576..8779
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8730..8750
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8824..8843
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8824..8842
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      join(8896..8901,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggagctgt atttcggcga ataccaacac gtacagcaag aatatggcgt ccatctgaga
       61 ctcgcaagcg atgatagcaa aaagccaagg aattcccaac actctaaggc aggctcctat
      121 ggtgtcagta ttcgggtgca gggaatcgat ggtcacccct acatagtcct gaataacaca
      181 gagcggtgtc tggcaggcac atcttttcct gaaaatgggc caccatttcc atctccaatg
      241 agaagtaacc tgcctctgca ttccagcaac gggtccgtgc tggaggagag cagcgcagag
      301 gaattgcagc tcccagaaaa tccgtatgcc cagcccagcc cagtaagaaa cctgaaacag
      361 tcctctctct tcgaaggcaa gaacggggtt ctagaacgca aagaggggcc aatgaagcca
      421 tcccacgtgt tgaactttca gaggcatcca gagcttctgc agccctatga ccctgaaaag
      481 aatgaatcga acttgcaaaa ccaccagcct cctgagcata attggctaaa cacattgaca
      541 gaagaaggta ccaacaataa gaaggcatgg acctgcttcc ccaagcctgg tagttcccag
      601 cctaccagcc ctcctttgga agatgcgggc aaatccggcg tgacggccat ccgtttgtgc
      661 agctctgtgg tcatcgatga ccccaagaag cagacctcag tgtgcgtaaa tgttgagagc
      721 tgcaccgggg aggcgctggt aggggaggcc ccttccccta gtgggaggtc ccccactgcc
      781 cacagcccgc acacccatcc tgaagccaag aaagccaggc cggacgtgct tcccttccgg
      841 cgacaagatt cagcgggacc tgtcctggac ggagctcggt cccggaggtc ctcttcctcg
      901 tcggcaaccc ccacgtcggc caactctttg tacagatttc tgcttgatga tcaggaatgt
      961 gccatccacg ctgacaatgt gaatcgccac gaaaacagac ggtatatccc cttcctgcca
     1021 ggaactgggc gggatattga cacgggatca attcctggtg tggatcaact gattgaaaag
     1081 tttgaccaaa aacccgggct tcagaggaga ggaaggcctg ggaagcggaa cagaatcaat
     1141 ccagaagaca ggaagaggtc cagaagcgtg gatagtgcct ttcctttcgg tctgcagggg
     1201 aactcggagt acctcagcga atttagccgg aacctgggca agtccagcga acacctcctc
     1261 cgcccgtcgc aggtgtgccc gcagcggtca cctgcccagg agcaccgcgg gagacagagc
     1321 gtgggccgcg ccttcgccaa gctgcagggg gcccacctcc ggcctgccca gcagagcaaa
     1381 gatgggaaag ctccagcaaa cagagggggt cgggagagta caaacacgcg tgtgtcctcc
     1441 cccccagccc agaataaaaa ggaggacgaa atcaagacag ccacggcgac gctgatgtta
     1501 cagaatcggg ccctagcgac ttcgcccgac tctggtgcca agaagatttc tgtgaagaca
     1561 tttacttccg cctccaacac tcaggccaca ccagatctgt tgaaaggcca acaagagctc
     1621 actcaacaaa ccaatgagga gacggccaag cagatacttt acaattacct caaagaagga
     1681 agcaccgata atgatgatgc gactaaacgg aaagtcaacc tggtctttga gaaaatccag
     1741 actttaaagt ctcgagcagc agggagcaca caaggaaata atcaagcttc taattcttca
     1801 tctgaagtca gagatctgct ggaacagaaa aacaagctga ccacagaagt ggctgaactt
     1861 cagagacagc ttcaactaga agtcaagaat cagcagaaca tcaaagaaga gagagagaga
     1921 atgaaagcaa atttggagga gctccagggc cagcatgata gtaaagtgga agagaactcc
     1981 atgctacagc agcgcctgga agagagtgaa ggggagctcc gcaagaacct ggaggaattg
     2041 ttccaggtga agatggaacg ggaacagcac cagactgaga tcagggatct ccaggaccag
     2101 ctctcagaaa tgcacgatga attggacagt gctaaacgat ctgaggacag ggaaaaggga
     2161 gccctgattg aggaactctt acaggcaaaa caggatcttc aggatctgct catcgccaaa
     2221 gaggagcagg aggacctctt gaggaagcgg gagcgtgaac tgaccgccct gaagggagcc
     2281 ctgaaggaag aggtgtccag ccacgacctg gagatggata agctgaagga gcagtatgac
     2341 gccgagctgc aggccctgag ggagagtgta gaagaagcaa ccaagaacgt cgaggtcctg
     2401 gccagtcgga gcaacacttc agagcagagc cagctggggg ctgaaatgca cgtgaaggct
     2461 ctgcaggagg agaatgagaa gctgcgggaa agcatcgaag agctggagca gagcgtcgct
     2521 cggctgcaga ggcagattgc tgacatgaag gacgatgagg tcaaggcaaa ggaaaggctc
     2581 acaaaatgcg agagagaaaa ccggcaatta gaggaggccc ttgtgaatgc aagaaaggaa
     2641 gaaagagaag ccacatcagt cagaaggacc ctggagggtg agctggaaga cgtgcagaga
     2701 aatctgagtc gggccaccca ggagcagaag cagttgtctg agaagcttaa agatgagact
     2761 gagcagaaag agcagttaag gaggctgaag aacgagatgg agaacgagcg atggcatctg
     2821 gacaagacca ttgaaaaatt gcagaaagag atggctgaca ttgttgaggt gtcccgtgta
     2881 tcaacactgg agctccagaa ccagctagat gagtacaagg agaaaaatcg cagggaactt
     2941 gcagaaatgc aaaggcagtt gaaggagaaa acgctagagg ctgaaaagtc acgtctgact
     3001 gccatgaaac tgcaggatga gatgcgtctg atggaggagg agttacggga ctaccagaga
     3061 gctcaggatg aggcgctcac aaaaaggcag cttctggagc agacactcaa ggacctggaa
     3121 tacgagctgg aggccaaaag tcacctcaaa gatgaccgaa gcagactcgt caagcagatg
     3181 gaggacaagg tgtctcagct ggagatggaa ctggaagaag aaagaaataa ctcggatttg
     3241 ctctcggaga ggatcactcg gagcagggaa cagatggagc agatgaggag cgagctcctg
     3301 caggaaagag ctctcaagca ggacttggag tgtgacaaga tttccttgga gaggcagaac
     3361 aaagatttaa agagccggat tatccatctg gaaggctcct acaggtccag caaagagggg
     3421 ctggtggtgc agatggaggc caggatcgcg gagctggagg accgtctgga aaacgaggag
     3481 agggaccggg ccagcctcca gctcagcaac cgacggctgg agcggaaggt gaaggagcta
     3541 gtgatgcaag tggacgatga gcacctgtct ctgactgatc agaaggacca gcttagcttg
     3601 cgcttgaaag caatgaaacg acaagtagag gaggccgagg aggagatcga cagactagaa
     3661 agttctaaaa agaagctgca gagggagctg gaggagcaga tggatgtgaa tgagcagctg
     3721 cagggacaac tcaattccat gaagaaggac ctaagactca agaagctgcc gagcaaagtg
     3781 ctggacgaca cggatgatga tgacctcagt acggacgggg ggagcctcca cgaggcgccg
     3841 ctgagctaca ccttccccaa ggacagcgct gccaccagcc agatcgatga attctaccca
     3901 tacgacgtcc cagactacgc ttgagcggcc gctcgagtct agagggcccg tttaaacccg
     3961 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4021 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4081 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4141 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     4201 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4261 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4321 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4381 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4441 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4501 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4561 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4621 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4681 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4741 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4801 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4861 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4921 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4981 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5041 attttcggat ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag
     5101 tataatacga caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc
     5161 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     5221 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     5281 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     5341 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     5401 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     5461 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt
     5521 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5581 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5641 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5701 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5761 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7981 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     8041 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     8101 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     8161 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     8221 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8281 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8341 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8401 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8461 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8521 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     8581 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     8641 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     8701 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     8761 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     8821 aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttggt
     8881 accgagctcg gatccgccac c
//
