LOCUS       sequence-436898-        7451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human Sema4D with N terminal myc after signal peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7451)
  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 7451)
  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
FEATURES             Location/Qualifiers
     source          1..7451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          689..1705
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1629..1651
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1713..1732
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1824..1853
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4472..4491)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4537..4592
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4591..4610)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4953..4969)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4953..4969)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4966..4988)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4977..4993
                     /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(5001..5007,5008..5025,5026..5031))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5046..5067
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5126..5321
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5172..5307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5327..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5364..5383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5418..5437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5529..5546)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5700..6288)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5780..5799)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6459..7250,7251..7319))
                     /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     7082..7101
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7320..7424)
                     /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 cccgggcggg gcgggggggg gggggggggg
      541 gggggggggg ggggggggtg gggggcgggg cggggcgagg cggagaggtg cggcggcagc
      601 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
      661 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      721 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      781 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      841 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
      901 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
      961 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg
     1021 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga
     1081 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc
     1141 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc
     1201 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg
     1261 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga
     1321 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg
     1381 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga
     1441 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg
     1501 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct
     1561 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
     1621 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
     1681 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
     1741 tcattttggc aaagatgagg atgtgcaccc ccattagggg gctgctcatg gcccttgcag
     1801 tgatgtttgg gacagcgatg gcagaacaaa aactcatctc agaagaggat ctgtttgcac
     1861 ccataccccg gatcacctgg gagcacagag aggtgcacct ggtgcagttt catgagccag
     1921 acatctacaa ctactcagcc ttgctgctga gcgaggacaa ggacaccttg tacataggtg
     1981 cccgggaggc ggtcttcgct gtgaacgcac tcaacatctc cgagaagcag catgaggtgt
     2041 attggaaggt ctcagaagac aaaaaagcaa aatgtgcaga aaaggggaaa tcaaaacaga
     2101 cagagtgcct caactacatc cgggtgctgc agccactcag cgccacttcc ctttacgtgt
     2161 gtgggaccaa cgcattccag ccggcctgtg accacctgaa cttaacatcc tttaagtttc
     2221 tggggaaaaa tgaagatggc aaaggaagat gtccctttga cccagcacac agctacacat
     2281 ccgtcatggt tgatggagaa ctttattcgg ggacgtcgta taattttttg ggaagtgaac
     2341 ccatcatctc ccgaaattct tcccacagtc ctctgaggac agaatatgca atcccttggc
     2401 tgaacgagcc tagtttcgtg tttgctgacg tgatccgaaa aagcccagac agccccgacg
     2461 gcgaggatga cagggtctac ttcttcttca cggaggtgtc tgtggagtat gagtttgtgt
     2521 tcagggtgct gatcccacgg atagcaagag tgtgcaaggg ggaccagggc ggcctgagga
     2581 ccttgcagaa gaaatggacc tccttcctga aagcccgact catctgctcc cggccagaca
     2641 gcggcttggt cttcaatgtg ctgcgggatg tcttcgtgct caggtccccg ggcctgaagg
     2701 tgcctgtgtt ctatgcactc ttcaccccac agctgaacaa cgtggggctg tcggcagtgt
     2761 gcgcctacaa cctgtccaca gccgaggagg tcttctccca cgggaagtac atgcagagca
     2821 ccacagtgga gcagtcccac accaagtggg tgcgctataa tggcccggta cccaagccgc
     2881 ggcctggagc gtgcatcgac agcgaggcac gggccgccaa ctacaccagc tccttgaatt
     2941 tgccagacaa gacgctgcag ttcgttaaag accacccttt gatggatgac tcggtaaccc
     3001 caatagacaa caggcccagg ttaatcaaga aagatgtgaa ctacacccag atcgtggtgg
     3061 accggaccca ggccctggat gggactgtct atgatgtcat gtttgtcagc acagaccggg
     3121 gagctctgca caaagccatc agcctcgagc acgctgttca catcatcgag gagacccagc
     3181 tcttccagga ctttgagcca gtccagaccc tgctgctgtc ttcaaagaag ggcaacaggt
     3241 ttgtctatgc tggctctaac tcgggcgtgg tccaggcccc gctggccttc tgtgggaagc
     3301 acggcacctg cgaggactgt gtgctggcgc gggaccccta ctgcgcctgg agcccgccca
     3361 cagcgacctg cgtggctctg caccagaccg agagccccag caggggtttg attcaggaga
     3421 tgagcggcga tgcttctgtg tgcccggata aaagtaaagg aagttaccgg cagcattttt
     3481 tcaagcacgg tggcacagcg gaactgaaat gctcccaaaa atccaacctg gcccgggtct
     3541 tttggaagtt ccagaatggc gtgttgaagg ccgagagccc caagtacggt cttatgggca
     3601 gaaaaaactt gctcatcttc aacttgtcag aaggagacag tggggtgtac cagtgcctgt
     3661 cagaggagag ggttaagaac aaaacggtct tccaagtggt cgccaagcac gtcctggaag
     3721 tgaaggtggt tccaaagccc gtagtggccc ccaccttgtc agttgttcag acagaaggta
     3781 gtaggattgc caccaaagtg ttggtggcat ccacccaagg gtcttctccc ccaaccccag
     3841 ccgtgcaggc cacctcctcc ggggccatca cccttcctcc caagcctgcg cccaccggca
     3901 catcctgcga accaaagatc gtcatcaaca cggtccccca gctccactcg gagaaaacca
     3961 tgtatcttaa gtccagcgac aaccgcctcc tcatgtccct cttcctcttc ttctttgttc
     4021 tcttcctctg cctctttttc tacaactgct ataagggata cctgcccaga cagtgcttga
     4081 aattccgctc ggccctacta attgggaaga agaagcccaa gtcagatttc tgtgaccgtg
     4141 agcagagcct gaaggagacg ttagtagagc cagggagctt ctcccagcag aatggggagc
     4201 accccaagcc agccctggac accggctatg agaccgagca agacaccatc accagcaaag
     4261 tccccacgga tagggaggac tcacagagga tcgacgacct ttctgccagg gacaagccct
     4321 ttgacgtcaa gtgtgagctg aagttcgctg actcagacgc agatggagac tgacctcgag
     4381 gatatcgcgg ccgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     4441 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     4501 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     4561 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     4621 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     4681 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     4741 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     4801 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     4861 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     4921 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     4981 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5041 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5101 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     5161 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5221 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5281 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     5341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     5461 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5521 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5581 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5641 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5701 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5761 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5821 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5881 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5941 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6001 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6061 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6421 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6481 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6541 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6601 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6661 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6721 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6781 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6841 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6901 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6961 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7021 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7081 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7141 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7201 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7261 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7321 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7381 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7441 gtgccacctg g
//
