LOCUS       sequence-436892-        6830 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Mouse Sema7A with SEMA domain deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6830)
  TITLE     Sema4D-Sema7A Interaction controls Migration and Axonal Acquisition 
            of Cortical Projection Neurons downstream of Satb2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6830)
  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            "Unpublished"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-Delta-SEMA
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(2462..3036,3037..3039,3040..3048)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2629..2646)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2856..2875
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3049..3051,3052..3054,3055..3768)
                     /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(3049..3051,3052..3054,3055..3765)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3094..3115)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3355..3374)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3702..3723
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3852..3871)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3917..3972
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3971..3990)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4333..4349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4333..4349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4346..4368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4357..4373
                     /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(4381..4387,4388..4405,4406..4411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4426..4447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4506..4701
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4552..4687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4707..4841
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4744..4763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4798..4817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4909..4926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5080..5668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5160..5179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5839..6630,6631..6699))
                     /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     6462..6481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6700..6804)
                     /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 gggggsgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tctgcgggat
     1741 gacgcctcct cctcccggac gtgccgcccc cagcgcaccg cgcgcccgcg tcctcagcct
     1801 gccggctcgg ttcgggctcc cgctgcggct gcggcttctg ctggtgttct gggtggccgc
     1861 cgcctccgcc caaggccact cgaggagcgg accccgcctg gacatgtgtg aggtctacag
     1921 cgggggctgc catggctgcc tcatgtcccg agacccctac tgtggctggg accaggaccg
     1981 ctgcgtgtct atctacagct cccaacgatc agtgctgcag tccattaatc cagcggagcc
     2041 acacagagag tgtcccaacc ctaaaccaga cgaggctcca ttgcagaagg tttccctggc
     2101 tcggaactct cgatactacc tgacctgccc catggagtcc cgccacgcca cttacttatg
     2161 gcgccatgag gagaatgtgg aacagagctg tgagccaggc caccaaagcc ctagctgcat
     2221 cctgttcatt gagaacctca cggcccgtca gtatggccac taccgctgcg aggcccaaga
     2281 gggctcctac ctccgtgagg ctcaacactg ggagctgctg ccagaggaca gagcactggc
     2341 cgaacaacta atgggccatg cccgggccct ggccgcctcc ttctggctgg gggtcctgcc
     2401 cacactcata cttggtctgc tggttcacta ggcggccgct acgtaaattc cgcccccccc
     2461 ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     2521 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     2581 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     2641 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     2701 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     2761 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     2821 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     2881 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     2941 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     3001 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     3061 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3121 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     3181 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     3241 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     3301 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     3361 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     3421 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     3481 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     3541 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     3601 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     3661 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     3721 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccggccg
     3781 ccagaacagg cggccggccg ccagcacagt ggtcgatcga ccaatgccct ggctcacaaa
     3841 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     3901 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     3961 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     4021 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     4081 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     4141 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4201 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4261 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt
     4321 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4381 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4441 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     4501 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4621 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4681 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     4741 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4801 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     4861 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4921 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4981 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5041 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5101 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5161 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5221 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5281 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5341 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5401 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5461 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5521 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5581 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5641 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5701 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5761 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5821 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5881 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5941 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6001 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6061 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6121 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6181 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6241 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6301 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6361 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6421 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6481 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6541 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6601 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6661 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6721 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6781 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
