LOCUS       sequence-471089-        7643 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 7643)
  AUTHORS   Flinois A, Mutero-Maeda A, Montessuit S, Citi S
  TITLE     Evidence for an interaction of paracingulin with microtubules.
  JOURNAL   MicroPubl Biol. 2024 Oct 14;2024:10.17912/micropub.biology.001341. 
            doi: 10.17912/micropub.biology.001341. eCollection 2024.
  PUBMED    39469042
REFERENCE   2  (bases 1 to 7643)
  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: "MicroPubl 
            Biol."; date: "2024-10-14"; volume: 
            "2024:10.17912/micropub.biology.001341. doi"; pages: " 
            10.17912/micropub.biology.001341. eCollection 2024"
COMMENT     Sequence Label: Strep-His-Tev-GFP-hCGNL1
FEATURES             Location/Qualifiers
     source          1..7643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4780..4914
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4817..4836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4871..4890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5035..5200
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5390..5978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5879..5898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  6113..6336
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6403..6936)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7125..7130,7131..7147,7148..7153))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(7224..7257)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        7334..7425
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     7350..7367
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(7467..7472,7473..7496,7497..7532,7533..7556)
                     /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             7569..7598
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             7614..7634
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt
      721 accatggaac tgtacttcgg cgagtaccag cacgtgcagc aagagtacgg agtccatctc
      781 cgtttggcat ctgacgatac tcagaaatca aggtccagcc aaaactcaaa ggccggcagt
      841 tacggagtct cgatccgtgt tcagggtatt gacggccacc catacatcgt gctgaacaac
      901 actgagaggt gcctcgctgg aacaagcttc tctgaaaacg gtcctccctt cccaccgcct
      961 gtcattaaca acctgccgct ccattcttca aacggcagtg ttcctaagga aaactcggag
     1021 gaattgcagc tgcccgagaa cccatacgcc caaccgtccc ctatccgtaa cctgaaacag
     1081 cctctgctcc acgagggaaa gaacggtgtc ctcgatagaa aagacggatc cgttaagccg
     1141 agccacttgc tgaacttcca gcgccatcct gagctcttgc aaccgtacga ccctgaaaag
     1201 aacgagctca acttgcaaaa ccatcagccc tcagaaagta actggctgaa gaccctcact
     1261 gaggaaggaa tcaacaacaa gaaaccatgg acttgcttcc ccaagccatc gaactcccag
     1321 cccacaagcc catctttgga ggatcctgca aagtccggtg tcactgcgat ccgtctctgt
     1381 agttcggtgg tcattgagga ccccaagaaa cagacgagcg tctgcgttaa cgtgcaaggc
     1441 tgtaccaagg aaagagtggg agaggaagcc ttgttcactt ctggaaggcc tctgacagca
     1501 cacagcccgc acgcgcatcc tgagacgaag aaaaccaggc ctgacgttct cccaatcagg
     1561 aggcaggata gtgctggtcc tgtgttggac ggagctaggt ctaggagatc cagctcttca
     1621 agtaccactc cgacccctgc taactccctg tacaggttcc tgctcgacga tcaggagtgt
     1681 gccatccacg cagacaacgt gaaccgtcat gaaaaccgcc gttacattcc gttcctgcct
     1741 ggaacgggta gggatatcga caccggcagc attcccggag tcgatcagct catcgagaaa
     1801 ttcgaccaga agccaggttt gcaaaggaga ggccgttctg gaaagcgcaa ccgtattaac
     1861 ccggacgata ggaagagatc acgcagtgtc gactcagctt tccctttcgg tctccagggc
     1921 aactccgaat acttgatcga gttcagcagg aacctgggca agtcgtccga gcacttgctg
     1981 cgtccttctc aggtgtgccc acaaaggcca ctgtctcagg aacgccgtgg taaacaatca
     2041 gtcggcagaa ccttcgctaa gctccaaggt gctgcacacg gagcttcttg tgctcattca
     2101 cgccctcctc agccaaacat cgacggcaag gtcctggaga cggaaggcag ccaggagtct
     2161 accgttattc gtgctccttc tctcggtgcc caatcaaaga aagaggaaga ggttaagaca
     2221 gccacggcaa ccctgatgct ccagaacagg gcagcggcta cgtcaccaga tagtggagct
     2281 aagaaaatct ctgtgaagac cttcccctcg gcatccaaca ctcaggcgac accagacctc
     2341 ttgaagggtc agcaagagct gacgcagcaa accaacgaag agaccgccaa acaaatcttg
     2401 tacaactacc tgaaggaggg ctcaactgac aacgacgatg ctacaaaaag aaaggtgaac
     2461 ttggtcttcg aaaaaattca gactctgaag agtcgcgccg caggatcggc tcagggtaac
     2521 aaccaagcct gcaacagcac ttctgaagtg aaggacctgc tcgagcaaaa atctaagctc
     2581 acaatcgaag tggctgagtt gcagagacaa ttgcagctgg aggtcaaaaa ccagcaaaac
     2641 attaaggaag agcgtgaaag gatgagagcc aacttggaag agctgcgctc acagcacaac
     2701 gagaaggtcg aagagaactc cacgctccag caacgtttgg aagagagcga gggagaactg
     2761 aggaaaaacc tcgaagagtt gttccaagtg aagatggagc gtgaacaaca ccagacagag
     2821 atcagggatc tgcaagacca gctctctgaa atgcatgatg agctggacag tgctaaaaga
     2881 tcggaagatc gcgagaaggg tgcactgatc gaagagttgc tgcaagcgaa gcaggatctc
     2941 caagacctct tgattgctaa ggaaggccgt gaggacctgc tccgcaagcg tgagagggaa
     3001 ctgacagcac tcaaaggagc gctgaaggaa gaggtcagct ctcacgatca ggagatggac
     3061 aaactcaagg aacagtacga tgctgagttg caagccctgc gcgagtccgt tgaagaggct
     3121 accaagaacg tcgaagttct ggcctcacgt agtaacacta gcgaacaaga ccaggccggt
     3181 acagagatga gggttaaatt gctgcaggaa gagaacgaga agctgcaagg cagatccgaa
     3241 gagctcgaaa ggagagtggc tcaattgcag cgccaaatcg aggatctgaa gggtgacgaa
     3301 gcgaaagcta aggagactct gaagaaatac gagggcgaaa ttagacagtt ggaagaggca
     3361 ctggtgcacg cgcgcaaaga agagaaggaa gccgtctctg cacgccgtgc gctcgagaac
     3421 gaattggagg gagctcaggg taacctgtca caaacaacgc aggaacaaaa gcagctgtct
     3481 gagaaactca aggaagagtc agaacagaaa gagcaactca ggagattgaa gaacgaaatg
     3541 gagaacgaaa gatggcatct cggcaagacc atcgaaaaat tgcagaagga gatggctgat
     3601 attgtcgaag cctcgcgcac ttccacactc gagttgcaaa accagttgga cgaatacaaa
     3661 gagaagaacc gccgtgaact ggccgagatg caacgtcagc tcaaagagaa gactttggag
     3721 gctgaaaaat cccgtctgac agccatgaag atgcaggacg gaatgaggct gatggaagag
     3781 gaactccgtg attaccagag ggctcaagac gaagccctga cgaagcgcca gctcttggag
     3841 caaaccctga aggacctcga gtacgaactc gaggctaaaa gtcacttgaa ggacgataga
     3901 tcgcgcctgg ttaaacagat ggaggctaag gtgtcccaat tggaaatgga gctggaggaa
     3961 gagagaaaca actccgacct gctcagcgag cgcatctcgc gttccaggga acagatggag
     4021 caactgagaa acggattgct gcaggaacgt gcggctaggc aagatctgga atgcggcaag
     4081 attagcctcg agagacagaa caaagacttg aagagtcgca tcattcacct ggagggatcg
     4141 tacaggtcta gtaaggaagg tctggttgtg caaatggagg ctcgtatcgc cgaactcgag
     4201 gataggttgg aaagtgaaga gagagaccgc gctaacctgc agctctccaa caggagactc
     4261 gaacgtaaag tcaaggagtt ggttatgcaa gtggacgatg agcacttgtc tctgactgat
     4321 cagaaggacc aactctcatt gagactgaaa gcaatgaagc gccaggtgga agaggcggaa
     4381 gaggaaatcg accgtctgga atcgtccaag aaaaagctgc agagggagct cgaggaacaa
     4441 atggatatga acgaacattt gcagggtcag ctgaactcaa tgaaaaagga cctccgcttg
     4501 aaaaagttgc ccagtaaggt gctggacgat atggacgatg acgatgacct gtccacagat
     4561 ggtggcagcc tctacgaggc tccagttagc tacacgttca gcaaggactc taccgtggcc
     4621 tctcagatct ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata
     4681 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     4741 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     4801 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4861 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatctgatca
     4921 ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     4981 tgccaccgct gagcaataac tatcataacc cctagggtat acccatctaa ttggaaccag
     5041 ataagtgaaa tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac
     5101 gctacaccca gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat
     5161 ttccacccct cccagttccc aactattttg tccgcccaca accggttgac ttgggtcaac
     5221 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5281 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5341 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5401 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5461 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5521 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5581 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5641 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5701 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5761 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5821 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5881 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5941 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6001 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6061 attgacttgg gtcgctcttc ctgtggatgc gcagatgccc tgcgtaagcg ggtgtgggcg
     6121 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     6181 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     6241 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     6301 gaggggcgtg gccaagggca tgtaaagact atattcgcgg cgttgtgaca atttaccgaa
     6361 caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg
     6421 gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc
     6481 gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc
     6541 atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac
     6601 tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca gaacgtaagc
     6661 cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact
     6721 acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc
     6781 tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc
     6841 gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact
     6901 gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac
     6961 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa
     7021 aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca
     7081 aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac
     7141 aggcttatgt caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa
     7201 ccttgggcag cagcgaagtc gccataactt cgtatagcat acattatacg aagttatctg
     7261 taactataac ggtcctaagg tagcgagttt aaacactagt atcgattcgc gacctactcc
     7321 ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta
     7381 ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca
     7441 taccgtccca ccatcgggcg cggatgtctg cttggtcaca ccctcaattc gagaaaggcg
     7501 gcggtagtgg cggcggatct ggcggtagtg cctggagcca tccgcagttt gaaaaggccg
     7561 gccatatgca ccaccatcat catcaccatc accatcacgg atctggcggt agtgaaaacc
     7621 tgtattttca gggcgccggt acc
//
