LOCUS       sequence-436848-        7424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mouse Sema4D with C Terminal Flag Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  AUTHORS   Bessa P, Newman AG, Yan K, Schaub T, Dannenberg R, Lajko D, 
            Eilenberger J, Brunet T, Textoris-Taube K, Kemmler E, Deng P, 
            Banerjee P, Ravindran E, Preissner R, Rosario M, Tarabykin V
  TITLE     Semaphorin heterodimerization in cis regulates membrane targeting 
            and neocortical wiring.
  JOURNAL   Nat Commun. 2024 Aug 16;15(1):7059. doi: 10.1038/s41467-024-51009-1.
  PUBMED    39152101
REFERENCE   2  (bases 1 to 7424)
  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: "Nat 
            Commun."; date: "2024-08-16"; volume: "15(1):7059. doi"; pages: " 
            10.1038/s41467-024-51009-1"
COMMENT     Sequence Label: pCAG-Sema4D-Flag
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..663
                     /label=chicken beta-actin promoter
     intron          664..1680
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1604..1626
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1688..1707
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             4334..4357
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4446..4465)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4511..4566
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4565..4584)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4927..4943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4927..4943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4940..4962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4951..4967
                     /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(4975..4981,4982..4999,5000..5005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5020..5041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5100..5295
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5146..5281
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5301..5435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5338..5357)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5392..5411
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5503..5520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5674..6262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5754..5773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6433..7224,7225..7293))
                     /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     7056..7075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7294..7398)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
      541 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
      601 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
      661 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
      721 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
      781 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
      841 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
      961 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     1021 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     1081 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     1141 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     1201 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     1261 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     1321 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     1381 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     1441 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     1501 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     1561 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     1621 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     1681 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcacccgg
     1741 gctagccacc atgaggatgt gtgcccccgt tagggggctg ttcttggccc tggtggtagt
     1801 gttgagaacc gcggtggcat ttgcacctgt gcctcggctc acctgggaac atggagaggt
     1861 aggtctggtg cagtttcaca agccaggcat ctttaactac tcggccttgc tgatgagtga
     1921 ggacaaagac actctgtatg taggcgcccg ggaagcagtc tttgcagtga atgcgctgaa
     1981 catctctgag aagcaacatg aggtatattg gaaggtctct gaagacaaaa aatccaagtg
     2041 tgcagagaag gggaaatcaa agcagacgga atgcctaaac tacattcgag tactacagcc
     2101 actaagcagc acttccctct atgtgtgtgg gaccaatgcg ttccagccca cctgtgacca
     2161 cctgaacttg acatccttca agtttctggg gaaaagtgaa gatggcaaag gaagatgccc
     2221 cttcgacccc gcccacagct acacatcagt catggttggg ggcgagctct actctgggac
     2281 gtcctataat ttcttgggca gtgaacccat catctctcga aactcttccc acagtccctt
     2341 gaggacggag tatgccatcc cgtggctgaa cgagcctagc ttcgtctttg ctgacgtgat
     2401 ccagaaaagc ccagatggtc cggagggtga agatgacaag gtctacttct tttttacgga
     2461 ggtatccgtg gagtacgaat tcgtcttcaa gttgatgatc ccgcgagttg ccagggtgtg
     2521 caagggcgac cagggcggcc tgcggacttt gcaaaaaaag tggacctcct tcctaaaggc
     2581 caggctgatc tgctccaagc cagacagtgg cctggtcttc aacatacttc aggatgtgtt
     2641 tgtgctgagg gccccgggcc tcaaggagcc tgtgttctat gcggtcttca ccccacagct
     2701 gaacaatgtg ggtctgtcag cggtgtgcgc ctacacactg gccacggtgg aggcagtctt
     2761 ctcccgtgga aagtacatgc agagtgccac agtggagcag tctcacacca agtgggtgcg
     2821 ctacaatggc ccagtgccca ctccccgacc tggagcgtgt atcgacagtg aggcccgggc
     2881 agccaactac accagctcct tgaatctccc agacaaaaca ctgcagtttg taaaagacca
     2941 ccctttgatg gatgactcag tgaccccgat agacaacaga cccaagctga tcaaaaaaga
     3001 tgtaaactac acccagatag tggtagacag gacccaggcc ctggatggga ctttctacga
     3061 cgtcatgttc atcagcacag accggggagc tctgcataaa gcagtcatcc tcacaaaaga
     3121 ggtgcatgtc atcgaggaga cccaactctt ccgggactct gaaccggtcc taactctgct
     3181 gctatcgtca aagaagggga ggaagtttgt ctatgcaggc tccaactctg gagtggtcca
     3241 agcgcccctg gcattctgcg aaaagcacgg tagctgtgaa gactgtgtgt tagcacggga
     3301 cccctactgt gcctggagcc cagccatcaa ggcctgtgtt accctgcacc aggaagaggc
     3361 ctccagcagg ggctggattc aggacatgag cggtgacaca tcctcatgcc tggataagag
     3421 taaagaaagt ttcaaccagc attttttcaa gcacggcggc acagcggaac tcaaatgttt
     3481 ccaaaagtcc aacctagccc gggtggtatg gaagttccag aatggcgagt tgaaggccgc
     3541 aagtcccaag tacggctttg tgggcaggaa gcacctgctc atcttcaacc tgtcggacgg
     3601 agacagcggc gtgtaccagt gcctgtcaga ggaaagggtg aggaataaaa cggtctccca
     3661 gctgctggcc aagcacgttc tggaagtgaa gatggtacct cggacccccc cctcacctac
     3721 ctcagaggat gctcagacag aaggtagtaa gatcacatcc aaaatgccgg ttgcatctac
     3781 ccaggggtcc tctcccccta ccccggctct gtgggcaacc tcccccagag ccgccaccct
     3841 acctcccaag tcctcctccg gcacatcctg tgaaccaaag atggtcatca acacggtccc
     3901 ccagctccac tcagagaaga cggtgtatct caagtccagt gacaaccgcc tgctcatgtc
     3961 tctcctcctc ttcatctttg tcctcttcct ctgcctcttt tcctacaact gctacaaggg
     4021 ctacctgccc ggacagtgct taaaattccg ctcagccctg ctgcttggaa agaaaacacc
     4081 caagtcagac ttctctgacc tggagcagag tgtgaaggag acactggtcg agcctgggag
     4141 cttctcccag cagaacggcg accaccccaa gccagccctg gatacgggct atgaaacgga
     4201 gcaggacacc atcaccagca aagtccccac ggatcgtgag gactcgcaac ggatcgatga
     4261 actctctgcc cgggacaaac cgtttgatgt caagtgtgaa ctgaagtttg cagattcgga
     4321 tgctgacggg gacgattata aagatgatga tgataaatga gcggccgcac tcctcaggtg
     4381 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     4441 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     4501 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     4561 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     4621 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     4681 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     4741 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     4801 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     4861 gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa gcttggcgta
     4921 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4981 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5041 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg
     5101 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5161 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5221 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5281 ctaggctttt gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca
     5341 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5401 ccaaactcat caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg
     5461 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5521 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5581 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctg
//
