LOCUS       sequence-436841-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Mouse Sema4D with N-terminal myc after endogenous Signal 
            peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  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 7430)
  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-sp-myc-mSema4D
FEATURES             Location/Qualifiers
     source          1..7430
                     /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             1820..1849
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4452..4471)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4517..4572
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4571..4590)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4933..4949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4933..4949)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4946..4968)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4957..4973
                     /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(4981..4987,4988..5005,5006..5011))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5026..5047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5106..5301
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5152..5287
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5307..5441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5344..5363)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5398..5417
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5509..5526)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5680..6268)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5760..5779)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6439..7230,7231..7299))
                     /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     7062..7081
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7300..7404)
                     /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 gcggtggcag aacagaaact gattagcgaa gaagatctgt ttgcacctgt
     1861 gcctcggctc acctgggaac atggagaggt aggtctggtg cagtttcaca agccaggcat
     1921 ctttaactac tcggccttgc tgatgagtga ggacaaagac actctgtatg taggcgcccg
     1981 ggaagcagtc tttgcagtga atgcgctgaa catctctgag aagcaacatg aggtatattg
     2041 gaaggtctct gaagacaaaa aatccaagtg tgcagagaag gggaaatcaa agcagacgga
     2101 atgcctaaac tacattcgag tactacagcc actaagcagc acttccctct atgtgtgtgg
     2161 gaccaatgcg ttccagccca cctgtgacca cctgaacttg acatccttca agtttctggg
     2221 gaaaagtgaa gatggcaaag gaagatgccc cttcgacccc gcccacagct acacatcagt
     2281 catggttggg ggcgagctct actctgggac gtcctataat ttcttgggca gtgaacccat
     2341 catctctcga aactcttccc acagtccctt gaggacggag tatgccatcc cgtggctgaa
     2401 cgagcctagc ttcgtctttg ctgacgtgat ccagaaaagc ccagatggtc cggagggtga
     2461 agatgacaag gtctacttct tttttacgga ggtatccgtg gagtacgaat tcgtcttcaa
     2521 gttgatgatc ccgcgagttg ccagggtgtg caagggcgac cagggcggcc tgcggacttt
     2581 gcaaaaaaag tggacctcct tcctaaaggc caggctgatc tgctccaagc cagacagtgg
     2641 cctggtcttc aacatacttc aggatgtgtt tgtgctgagg gccccgggcc tcaaggagcc
     2701 tgtgttctat gcggtcttca ccccacagct gaacaatgtg ggtctgtcag cggtgtgcgc
     2761 ctacacactg gccacggtgg aggcagtctt ctcccgtgga aagtacatgc agagtgccac
     2821 agtggagcag tctcacacca agtgggtgcg ctacaatggc ccagtgccca ctccccgacc
     2881 tggagcgtgt atcgacagtg aggcccgggc agccaactac accagctcct tgaatctccc
     2941 agacaaaaca ctgcagtttg taaaagacca ccctttgatg gatgactcag tgaccccgat
     3001 agacaacaga cccaagctga tcaaaaaaga tgtaaactac acccagatag tggtagacag
     3061 gacccaggcc ctggatggga ctttctacga cgtcatgttc atcagcacag accggggagc
     3121 tctgcataaa gcagtcatcc tcacaaaaga ggtgcatgtc atcgaggaga cccaactctt
     3181 ccgggactct gaaccggtcc taactctgct gctatcgtca aagaagggga ggaagtttgt
     3241 ctatgcaggc tccaactctg gagtggtcca agcgcccctg gcattctgcg aaaagcacgg
     3301 tagctgtgaa gactgtgtgt tagcacggga cccctactgt gcctggagcc cagccatcaa
     3361 ggcctgtgtt accctgcacc aggaagaggc ctccagcagg ggctggattc aggacatgag
     3421 cggtgacaca tcctcatgcc tggataagag taaagaaagt ttcaaccagc attttttcaa
     3481 gcacggcggc acagcggaac tcaaatgttt ccaaaagtcc aacctagccc gggtggtatg
     3541 gaagttccag aatggcgagt tgaaggccgc aagtcccaag tacggctttg tgggcaggaa
     3601 gcacctgctc atcttcaacc tgtcggacgg agacagcggc gtgtaccagt gcctgtcaga
     3661 ggaaagggtg aggaataaaa cggtctccca gctgctggcc aagcacgttc tggaagtgaa
     3721 gatggtacct cggacccccc cctcacctac ctcagaggat gctcagacag aaggtagtaa
     3781 gatcacatcc aaaatgccgg ttgcatctac ccaggggtcc tctcccccta ccccggctct
     3841 gtgggcaacc tcccccagag ccgccaccct acctcccaag tcctcctccg gcacatcctg
     3901 tgaaccaaag atggtcatca acacggtccc ccagctccac tcagagaaga cggtgtatct
     3961 caagtccagt gacaaccgcc tgctcatgtc tctcctcctc ttcatctttg tcctcttcct
     4021 ctgcctcttt tcctacaact gctacaaggg ctacctgccc ggacagtgct taaaattccg
     4081 ctcagccctg ctgcttggaa agaaaacacc caagtcagac ttctctgacc tggagcagag
     4141 tgtgaaggag acactggtcg agcctgggag cttctcccag cagaacggcg accaccccaa
     4201 gccagccctg gatacgggct atgaaacgga gcaggacacc atcaccagca aagtccccac
     4261 ggatcgtgag gactcgcaac ggatcgatga actctctgcc cgggacaaac cgtttgatgt
     4321 caagtgtgaa ctgaagtttg cagattcgga tgctgacggg gactgagcgg ccgcactcct
     4381 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     4441 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     4501 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     4561 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4621 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4681 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4741 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4801 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4861 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt
     4921 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4981 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     5041 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     5101 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5161 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5221 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     5281 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     5341 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5401 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     5461 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5521 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5581 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5641 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5701 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5761 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5821 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5881 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5941 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6001 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6061 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6121 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6181 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6241 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6301 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6361 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6421 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6481 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6541 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6601 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6661 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6721 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6781 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6841 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6901 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6961 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     7021 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7081 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7141 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7201 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7261 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7321 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7381 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
