LOCUS       sequence-436894-        7856 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Mouse Sema7A with IG Domain Deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7856)
  TITLE     Sema4D-Sema7A Interaction controls Migration and Axonal Acquisition 
            of Cortical Projection Neurons downstream of Satb2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7856)
  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            "Unpublished"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-IG
FEATURES             Location/Qualifiers
     source          1..7856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..662
                     /label=chicken beta-actin promoter
     intron          663..1679
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1603..1625
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1687..1706
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(3507..4081,4082..4084,4085..4093)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3674..3691)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3901..3920
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4094..4096,4097..4099,4100..4813)
                     /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(4094..4096,4097..4099,4100..4810)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4139..4160)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4400..4419)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4747..4768
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4877..4896)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4942..4997
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4996..5015)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5358..5374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5358..5374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5371..5393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5382..5398
                     /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(5406..5412,5413..5430,5431..5436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5451..5472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5531..5726
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5577..5712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5732..5866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5769..5788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5823..5842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5934..5951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6105..6693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6185..6204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6864..7655,7656..7724))
                     /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     7487..7506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7725..7829)
                     /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 ggggggcgcg cgccgggcgg ggcggggcgg ggcggggggc
      541 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      601 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      661 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
      841 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
      901 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1021 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1081 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1141 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1201 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1261 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1321 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1381 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1441 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1501 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1561 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1621 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1681 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttctgcggga
     1741 tgacgcctcc tcctcccgga cgtgccgccc ccagcgcacc gcgcgcccgc gtcctcagcc
     1801 tgccggctcg gttcgggctc ccgctgcggc tgcggcttct gctggtgttc tgggtggccg
     1861 ccgcctccgc ccaaggccac tcgaggagcg gaccccgcat ctccgccgtc tggaaagggc
     1921 aggaccatgt ggactttagc cagcctgagc cacacaccgt gcttttccat gagccgggca
     1981 gcttctctgt ctgggtgggt ggacgtggca aggtctacca cttcaacttc cccgagggca
     2041 agaatgcctc tgtgcgcacg gtgaacatcg gctccacaaa ggggtcctgt caggacaaac
     2101 aggactgtgg gaattacatc actcttctag aaaggcgggg taatgggctg ctggtctgtg
     2161 gcaccaatgc ccggaagccc agctgctgga acttggtgaa tgacagtgtg gtgatgtcac
     2221 ttggtgagat gaaaggctat gcccccttca gcccggatga gaactccctg gttctgtttg
     2281 aaggagatga agtgtactct accatccgga agcaggaata caacgggaag atccctcggt
     2341 ttcgacgcat tcggggcgag agtgaactgt acacaagtga tacagtcatg cagaacccac
     2401 agttcatcaa ggccaccatt gtgcaccaag accaagccta tgatgataag atctactact
     2461 tcttccgaga agacaaccct gacaagaacc ccgaggctcc tctcaatgtg tcccgagtag
     2521 cccagttgtg caggggggac cagggtggtg agagttcgtt gtctgtctcc aagtggaaca
     2581 ccttcctgaa agccatgttg gtctgcagcg atgcagccac caacaggaac ttcaatcggc
     2641 tgcaagatgt cttcctgctc cctgacccca gtggccagtg gagagatacc agggtctatg
     2701 gcgttttctc caacccctgg aactactcag ctgtctgcgt gtattcgctt ggtgacattg
     2761 acagagtctt ccgtacctca tcgctcaaag gctaccacat gggccttccc aaccctcgac
     2821 ctggcatgtg cctcccaaaa aagcagccca tacccacaga aaccttccag gtagctgata
     2881 gtcacccaga ggtggctcag agggtggaac ctatggggcc actgaagaca ccattgttcc
     2941 attctaagta ccattaccag aaagtggtcg ttcaccgcat gcaagccagc aatggagaga
     3001 ccttccatgt gctttatcta accacagaca ggggcaccat tcacaaggtg gtggaatcag
     3061 gggaccagga ccatagcttt gtcttcaata tcatggagat ccagcccttt caccgtgcag
     3121 cggccatcca ggctatatca ttggatgctg accggcggaa gctctatgtg acctcccagt
     3181 gggaagtgag ccaggtaccc ctggacatgt gtgaggtcta cagcgggggc tgccatggct
     3241 gcctcatgtc ccgagacccc tactgtggct gggaccagga ccgctgcgtg tctatctaca
     3301 gctcccaacg atcagtgctg cagtccatta atccagcgga gccacacaga gagtgtccca
     3361 acctgccaga ggacagagca ctggccgaac aactaatggg ccatgcccgg gccctggccg
     3421 cctccttctg gctgggggtc ctgcccacac tcatacttgg tctgctggtt cactaggcgg
     3481 ccgctacgta aattccgccc cccccccccc tctccctccc ccccccctaa cgttactggc
     3541 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     3601 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     3661 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     3721 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     3781 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     3841 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     3901 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     3961 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     4021 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     4081 tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     4141 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     4201 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4261 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4321 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     4381 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     4441 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     4501 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     4561 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     4621 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     4681 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     4741 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     4801 gctgtacaag taaagcggcc ggccgccagc acagtggtcg atcgaccaat gccctggctc
     4861 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     4921 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     4981 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     5041 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5101 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     5161 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     5221 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     5281 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     5341 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5521 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5581 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5641 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5701 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     5761 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5821 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     5881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6541 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7801 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgg
//
