LOCUS       sequence-231514-        6062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses RNAi-resistant Sema4A with N-terminal myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  AUTHORS   McDermott JE, Goldblatt D, Paradis S
  TITLE     Class 4 Semaphorins and Plexin-B receptors regulate GABAergic and 
            glutamatergic synapse development in the mammalian hippocampus.
  JOURNAL   Mol Cell Neurosci. 2018 Jul 4;92:50-66. doi: 
            10.1016/j.mcn.2018.06.008.
  PUBMED    29981480
REFERENCE   2  (bases 1 to 6062)
  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: "Mol Cell 
            Neurosci."; date: "2018-07-4"; volume: "92:50-66. doi"; pages: " 
            10.1016/j.mcn.2018.06.008"
COMMENT     Sequence Label: pCMV-myc-Sema4A RNAiR
FEATURES             Location/Qualifiers
     source          1..6062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      803..812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3175..3309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3199..3218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3253..3272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3420..3436)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3420..3436)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3433..3455)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3444..3460
                     /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(3468..3474,3475..3492,3493..3498))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3513..3534
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3651..3668)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3822..4410)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3902..3921)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4581..5372,5373..5441))
                     /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     5204..5223
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5442..5546)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5614..5632
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5670..5692)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5792..5811
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6005..6027
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6019..6036
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6020..6036
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctaaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggcccg ccgccaccat ggctgggatt ttctatttca tcctcttttc
      841 gtttctcttt ggaatttgcg acgctgtcac cggttctaga gggcccacca tggcatcaat
      901 gcagaagctg atctcagagg aggacctgct tatggccatg gaggcccgaa ctggtggtca
      961 ggggcccatg cccagagtca aataccatgc tggagacggg cgcagggccc tcagcttctt
     1021 ccaacaaaaa ggcctccgag actttgacac gctgctcctg agtgacgatg gcaacactct
     1081 ctatgtgggg gctcgagagg ccgtcctggc cttgaatatc cagaacccag gaatcccaag
     1141 gctaaagaac atgataccct ggccagccag tgagagaaaa aagaccgaat gtgcctttaa
     1201 gaagaagagc aatgagacac agtgtttcaa cttcattcga gtcctggtct cttacaatgc
     1261 tactcacctc tatgcctgtg ggacctttgc cttcagccct gcctgtacct tcattgaact
     1321 ccaagattcc ctcctgttgc ccatcttgat agacaaggtc atggacggga agggccaaag
     1381 cccctttgac cctgttcaca agcacacagc tgtcttggtc gatgggatgc tttattccgg
     1441 caccatgaac aacttcctgg gcagcgagcc catcctgatg cggacactgg gatcccagcc
     1501 tgttctcaag actgacatct tcttacgctg gctgcacgcg gatgcctcct tcgtggcagc
     1561 cattccatcc acccaggtcg tctatttctt ctttgaggag acagccagcg agtttgactt
     1621 ctttgaagag ctgtatatat ccagggtggc tcaagtctgc aagaacgacg tgggcggtga
     1681 aaagctactg cagaagaagt ggaccacctt cctcaaagcc cagttgctct gcgctcagcc
     1741 agggcagctg ccattcaaca tcatccgcca cgcggtcctg ctgcctgccg attctccctc
     1801 tgtttcccgc atctacgcag tctttacctc ccagtggcag gttggcggga ccaggagctc
     1861 agcagtctgt gccttctctc tcacggacat tgagcgagtc tttaaaggga agtacaagga
     1921 gctgaacaag gagacctccc gctggaccac ttgccggggc tcagaggtca gcccgaggcc
     1981 aggcagttgc tccatgggcc cctcctctga caaggcatta accttcatga aggaccattt
     2041 tctgatggat gagcacgtgg taggaacacc cctgctggtg aagtctggtg tggagtacac
     2101 acggcttgct gtggagtcag ctcggggcct tgatgggagc agccatgtgg tcatgtatct
     2161 gggtacctcc acggggtccc tgcacaaggc tgtggtgcct caggacagca gtgcttatct
     2221 cgtggaggag attcagctga gccctgactc tgagcctgtt cgaaacctgc agctggcccc
     2281 cgcccagggt gcagtgtttg caggcttctc tggaggcatc tggagagttc ccagggccaa
     2341 ttgcagtgtc tacgagagct gtgtggactg tgtgcttgcc agggaccctc actgtgcctg
     2401 ggaccctgaa tcaagactct gcagccttct gtctggctct accaagcctt ggaagcagga
     2461 catggaacgc ggcaacccgg agtgggtatg cacccgtggc cccatggcca ggagcccccg
     2521 gcgtcagagc ccccctcaac taattaaaga agtcctgaca gtccccaact ccatcctgga
     2581 gctgccctgc ccccacctgt cagcactggc ctcttaccac tggagtcatg gccgagccaa
     2641 aatctcagaa gcctctgcta ccgtctacaa tggctccctc ttgctgctgc cgcaggatgg
     2701 tgtcgggggc ctctaccagt gtgtggcgac tgagaacggc tactcatacc ctgtggtctc
     2761 ctattgggta gacagccagg accagcccct ggcgctggac cctgagctgg cgggcgttcc
     2821 ccgtgagcgt gtgcaggtcc cgctgaccag ggtcggaggc ggagcttcca tggctgccca
     2881 gcggtcctac tggccccatt ttctcatcgt taccgtcctc ctggccatcg tgctcctggg
     2941 agtgctcact ctcctcctcg cttccccact gggggcgctg cgggctcggg gtaaggttca
     3001 gggctgtggg atgctgcccc ccagggaaaa ggctccactg agcagggacc agcacctcca
     3061 gccctccaag gaccacagga cctctgccag tgacgtagat gccgacaaca accatctggg
     3121 cgccgaagtg gcttgacagt cgaccgagat ctctcgaggt accgcggccg cggggatcca
     3181 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     3241 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     3301 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg
     3361 gaggtttttt cggatcctct agagtcgatc tgcaggcatg ctagcttggc gtaatcatgg
     3421 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3481 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3541 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3601 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3661 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3721 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3781 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3841 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3901 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3961 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4021 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4081 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4141 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4201 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4261 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4321 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4381 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4441 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4501 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4561 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4621 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4681 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4741 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4801 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4861 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4921 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4981 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5041 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5101 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5161 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5221 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5281 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5341 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5401 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5461 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5521 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5581 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5641 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5701 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5761 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5821 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     5881 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     5941 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     6001 ttttcccagt cacgacgttg taaaacgacg gccagtgagt tcgagcttgc atgcctgcag
     6061 gt
//
