LOCUS       sequence-436901-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Sema4D with N terminal myc after signal peptide 
            containing 497P mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  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 7453)
  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-hSema4D-Q497P
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          691..1707
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1631..1653
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1715..1734
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1826..1855
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4474..4493)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4539..4594
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4593..4612)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4955..4971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4955..4971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4968..4990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4979..4995
                     /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(5003..5009,5010..5027,5028..5033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5128..5323
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5174..5309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5329..5463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5366..5385)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5420..5439
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5531..5548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5702..6290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5782..5801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6461..7252,7253..7321))
                     /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     7084..7103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7322..7426)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggggcg cctggggggg ggggtggggg gggggggggg
      541 gggggggggg gggggggggg gggggggcgg ggcggggcga ggcggagagg tgcggcggca
      601 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
      661 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
      721 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
      781 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
      841 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg
      901 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc
      961 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc
     1021 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc
     1081 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc
     1141 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg
     1201 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg
     1261 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg
     1321 gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat
     1381 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg
     1441 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc
     1501 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc
     1561 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg
     1621 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat
     1681 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct
     1741 catcattttg gcaaagatga ggatgtgcac ccccattagg gggctgctca tggcccttgc
     1801 agtgatgttt gggacagcga tggcagaaca aaaactcatc tcagaagagg atctgtttgc
     1861 acccataccc cggatcacct gggagcacag agaggtgcac ctggtgcagt ttcatgagcc
     1921 agacatctac aactactcag ccttgctgct gagcgaggac aaggacacct tgtacatagg
     1981 tgcccgggag gcggtcttcg ctgtgaacgc actcaacatc tccgagaagc agcatgaggt
     2041 gtattggaag gtctcagaag acaaaaaagc aaaatgtgca gaaaagggga aatcaaaaca
     2101 gacagagtgc ctcaactaca tccgggtgct gcagccactc agcgccactt ccctttacgt
     2161 gtgtgggacc aacgcattcc agccggcctg tgaccacctg aacttaacat cctttaagtt
     2221 tctggggaaa aatgaagatg gcaaaggaag atgtcccttt gacccagcac acagctacac
     2281 atccgtcatg gttgatggag aactttattc ggggacgtcg tataattttt tgggaagtga
     2341 acccatcatc tcccgaaatt cttcccacag tcctctgagg acagaatatg caatcccttg
     2401 gctgaacgag cctagtttcg tgtttgctga cgtgatccga aaaagcccag acagccccga
     2461 cggcgaggat gacagggtct acttcttctt cacggaggtg tctgtggagt atgagtttgt
     2521 gttcagggtg ctgatcccac ggatagcaag agtgtgcaag ggggaccagg gcggcctgag
     2581 gaccttgcag aagaaatgga cctccttcct gaaagcccga ctcatctgct cccggccaga
     2641 cagcggcttg gtcttcaatg tgctgcggga tgtcttcgtg ctcaggtccc cgggcctgaa
     2701 ggtgcctgtg ttctatgcac tcttcacccc acagctgaac aacgtggggc tgtcggcagt
     2761 gtgcgcctac aacctgtcca cagccgagga ggtcttctcc cacgggaagt acatgcagag
     2821 caccacagtg gagcagtccc acaccaagtg ggtgcgctat aatggcccgg tacccaagcc
     2881 gcggcctgga gcgtgcatcg acagcgaggc acgggccgcc aactacacca gctccttgaa
     2941 tttgccagac aagacgctgc agttcgttaa agaccaccct ttgatggatg actcggtaac
     3001 cccaatagac aacaggccca ggttaatcaa gaaagatgtg aactacaccc agatcgtggt
     3061 ggaccggacc caggccctgg atgggactgt ctatgatgtc atgtttgtca gcacagaccg
     3121 gggagctctg cacaaagcca tcagcctcga gcacgctgtt cacatcatcg aggagaccca
     3181 gctcttccag gactttgagc cagtccagac cctgctgctg tcttcaaaga agggcaacag
     3241 gtttgtctat gctggctcta actcgggcgt ggtccccgcc ccgctggcct tctgtgggaa
     3301 gcacggcacc tgcgaggact gtgtgctggc gcgggacccc tactgcgcct ggagcccgcc
     3361 cacagcgacc tgcgtggctc tgcaccagac cgagagcccc agcaggggtt tgattcagga
     3421 gatgagcggc gatgcttctg tgtgcccgga taaaagtaaa ggaagttacc ggcagcattt
     3481 tttcaagcac ggtggcacag cggaactgaa atgctcccaa aaatccaacc tggcccgggt
     3541 cttttggaag ttccagaatg gcgtgttgaa ggccgagagc cccaagtacg gtcttatggg
     3601 cagaaaaaac ttgctcatct tcaacttgtc agaaggagac agtggggtgt accagtgcct
     3661 gtcagaggag agggttaaga acaaaacggt cttccaagtg gtcgccaagc acgtcctgga
     3721 agtgaaggtg gttccaaagc ccgtagtggc ccccaccttg tcagttgttc agacagaagg
     3781 tagtaggatt gccaccaaag tgttggtggc atccacccaa gggtcttctc ccccaacccc
     3841 agccgtgcag gccacctcct ccggggccat cacccttcct cccaagcctg cgcccaccgg
     3901 cacatcctgc gaaccaaaga tcgtcatcaa cacggtcccc cagctccact cggagaaaac
     3961 catgtatctt aagtccagcg acaaccgcct cctcatgtcc ctcttcctct tcttctttgt
     4021 tctcttcctc tgcctctttt tctacaactg ctataaggga tacctgccca gacagtgctt
     4081 gaaattccgc tcggccctac taattgggaa gaagaagccc aagtcagatt tctgtgaccg
     4141 tgagcagagc ctgaaggaga cgttagtaga gccagggagc ttctcccagc agaatgggga
     4201 gcaccccaag ccagccctgg acaccggcta tgagaccgag caagacacca tcaccagcaa
     4261 agtccccacg gatagggagg actcacagag gatcgacgac ctttctgcca gggacaagcc
     4321 ctttgacgtc aagtgtgagc tgaagttcgc tgactcagac gcagatggag actgacctcg
     4381 aggatatcgc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     4441 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     4501 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     4561 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     4621 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     4681 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     4741 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     4801 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     4861 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     4921 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4981 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5041 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5101 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     5161 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5221 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5281 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     5341 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5401 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5461 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5521 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5581 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5641 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5701 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5761 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5821 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5881 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5941 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6001 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6061 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6121 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6181 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6241 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6301 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6361 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7441 aagtgccacc tgg
//
