LOCUS       sequence-436897-        8069 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Mouse Sema7A with RGD motif mutated to KCE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8069)
  TITLE     Sema4D-Sema7A Interaction controls Migration and Axonal Acquisition 
            of Cortical Projection Neurons downstream of Satb2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8069)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCAGIG-Sema7A-KCE
FEATURES             Location/Qualifiers
     source          1..8069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          620..1636
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1560..1582
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1644..1663
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3720..4294,4295..4297,4298..4306)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3887..3904)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4114..4133
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4307..4309,4310..4312,4313..5026)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4307..4309,4310..4312,4313..5023)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4352..4373)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4613..4632)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4960..4981
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5090..5109)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5155..5210
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5209..5228)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5571..5587)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5571..5587)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5584..5606)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5595..5611
                     /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(5619..5625,5626..5643,5644..5649))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5664..5685
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5744..5939
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5790..5925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5945..6079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5982..6001)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6036..6055
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6147..6164)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6318..6906)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6398..6417)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7077..7868,7869..7937))
                     /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     7700..7719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7938..8042)
                     /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 ggggggcgag gcggagaggt gcggcggcag ccaatcagag
      541 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      601 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
      661 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      721 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
      781 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
      841 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct
      901 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg
      961 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa
     1021 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg
     1081 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     1141 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     1201 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg
     1261 cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg
     1321 gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc
     1381 tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag
     1441 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct
     1501 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     1561 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt
     1621 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     1681 caaagaattc tgcgggatga cgcctcctcc tcccggacgt gccgccccca gcgcaccgcg
     1741 cgcccgcgtc ctcagcctgc cggctcggtt cgggctcccg ctgcggctgc ggcttctgct
     1801 ggtgttctgg gtggccgccg cctccgccca aggccactcg aggagcggac cccgcatctc
     1861 cgccgtctgg aaagggcagg accatgtgga ctttagccag cctgagccac acaccgtgct
     1921 tttccatgag ccgggcagct tctctgtctg ggtgggtgga cgtggcaagg tctaccactt
     1981 caacttcccc gagggcaaga atgcctctgt gcgcacggtg aacatcggct ccacaaaggg
     2041 gtcctgtcag gacaaacagg actgtgggaa ttacatcact cttctagaaa ggcggggtaa
     2101 tgggctgctg gtctgtggca ccaatgcccg gaagcccagc tgctggaact tggtgaatga
     2161 cagtgtggtg atgtcacttg gtgagatgaa aggctatgcc cccttcagcc cggatgagaa
     2221 ctccctggtt ctgtttgaag gagatgaagt gtactctacc atccggaagc aggaatacaa
     2281 cgggaagatc cctcggtttc gacgcattcg gggcgagagt gaactgtaca caagtgatac
     2341 agtcatgcag aacccacagt tcatcaaggc caccattgtg caccaagacc aagcctatga
     2401 tgataagatc tactacttct tccgagaaga caaccctgac aagaaccccg aggctcctct
     2461 caatgtgtcc cgagtagccc agttgtgcaa gtgcgagcag ggtggtgaga gttcgttgtc
     2521 tgtctccaag tggaacacct tcctgaaagc catgttggtc tgcagcgatg cagccaccaa
     2581 caggaacttc aatcggctgc aagatgtctt cctgctccct gaccccagtg gccagtggag
     2641 agataccagg gtctatggcg ttttctccaa cccctggaac tactcagctg tctgcgtgta
     2701 ttcgcttggt gacattgaca gagtcttccg tacctcatcg ctcaaaggct accacatggg
     2761 ccttcccaac cctcgacctg gcatgtgcct cccaaaaaag cagcccatac ccacagaaac
     2821 cttccaggta gctgatagtc acccagaggt ggctcagagg gtggaaccta tggggccact
     2881 gaagacacca ttgttccatt ctaagtacca ttaccagaaa gtggtcgttc accgcatgca
     2941 agccagcaat ggagagacct tccatgtgct ttatctaacc acagacaggg gcaccattca
     3001 caaggtggtg gaatcagggg accaggacca tagctttgtc ttcaatatca tggagatcca
     3061 gccctttcac cgtgcagcgg ccatccaggc tatatcattg gatgctgacc ggcggaagct
     3121 ctatgtgacc tcccagtggg aagtgagcca ggtacccctg gacatgtgtg aggtctacag
     3181 cgggggctgc catggctgcc tcatgtcccg agacccctac tgtggctggg accaggaccg
     3241 ctgcgtgtct atctacagct cccaacgatc agtgctgcag tccattaatc cagcggagcc
     3301 acacagagag tgtcccaacc ctaaaccaga cgaggctcca ttgcagaagg tttccctggc
     3361 tcggaactct cgatactacc tgacctgccc catggagtcc cgccacgcca cttacttatg
     3421 gcgccatgag gagaatgtgg aacagagctg tgagccaggc caccaaagcc ctagctgcat
     3481 cctgttcatt gagaacctca cggcccgtca gtatggccac taccgctgcg aggcccaaga
     3541 gggctcctac ctccgtgagg ctcaacactg ggagctgctg ccagaggaca gagcactggc
     3601 cgaacaacta atgggccatg cccgggccct ggccgcctcc ttctggctgg gggtcctgcc
     3661 cacactcata cttggtctgc tggttcacta ggcggccgct acgtaaattc cgcccccccc
     3721 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     3781 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     3841 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     3901 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     3961 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     4021 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     4081 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     4141 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     4201 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     4261 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     4321 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4381 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4441 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4501 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4561 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4621 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4681 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4741 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     4801 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4861 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4921 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4981 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccggccgcc
     5041 agcacagtgg tcgatcgacc aatgccctgg ctcacaaata ccactgagat ctttttccct
     5101 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     5161 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     5221 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     5281 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     5341 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     5401 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     5461 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5521 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     5581 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5641 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5701 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc aattagtcag
     5761 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5821 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5881 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5941 agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6001 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6061 atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6121 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6181 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6241 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6301 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6361 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6421 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6481 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6541 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6601 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6661 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6721 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6781 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6841 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6901 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6961 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7021 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7081 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7141 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7201 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7261 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7321 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7381 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     7441 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     7501 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7561 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7621 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7681 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7741 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7801 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7861 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7921 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7981 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8041 cgcacatttc cccgaaaagt gccacctgg
//
