LOCUS       sequence-436895-        7960 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Mouse Sema7A with PSI domain deleted.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7960)
  TITLE     Sema4D-Sema7A Interaction controls Migration and Axonal Acquisition 
            of Cortical Projection Neurons downstream of Satb2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7960)
  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            "Unpublished"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-PSI
FEATURES             Location/Qualifiers
     source          1..7960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..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(3611..4185,4186..4188,4189..4197)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3778..3795)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4005..4024
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4198..4200,4201..4203,4204..4917)
                     /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(4198..4200,4201..4203,4204..4914)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4243..4264)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4504..4523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4851..4872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4981..5000)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5046..5101
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5100..5119)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5462..5478)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5462..5478)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5475..5497)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5486..5502
                     /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(5510..5516,5517..5534,5535..5540))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5555..5576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5635..5830
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5681..5816
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5836..5970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5873..5892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5927..5946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6038..6055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6209..6797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6289..6308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6968..7759,7760..7828))
                     /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     7591..7610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7829..7933)
                     /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 gggggcgcgc gccaggcggg gcgggggggg 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 accccgcatc tccgccgtct ggaaagggca
     1921 ggaccatgtg gactttagcc agcctgagcc acacaccgtg cttttccatg agccgggcag
     1981 cttctctgtc tgggtgggtg gacgtggcaa ggtctaccac ttcaacttcc ccgagggcaa
     2041 gaatgcctct gtgcgcacgg tgaacatcgg ctccacaaag gggtcctgtc aggacaaaca
     2101 ggactgtggg aattacatca ctcttctaga aaggcggggt aatgggctgc tggtctgtgg
     2161 caccaatgcc cggaagccca gctgctggaa cttggtgaat gacagtgtgg tgatgtcact
     2221 tggtgagatg aaaggctatg cccccttcag cccggatgag aactccctgg ttctgtttga
     2281 aggagatgaa gtgtactcta ccatccggaa gcaggaatac aacgggaaga tccctcggtt
     2341 tcgacgcatt cggggcgaga gtgaactgta cacaagtgat acagtcatgc agaacccaca
     2401 gttcatcaag gccaccattg tgcaccaaga ccaagcctat gatgataaga tctactactt
     2461 cttccgagaa gacaaccctg acaagaaccc cgaggctcct ctcaatgtgt cccgagtagc
     2521 ccagttgtgc aggggggacc agggtggtga gagttcgttg tctgtctcca agtggaacac
     2581 cttcctgaaa gccatgttgg tctgcagcga tgcagccacc aacaggaact tcaatcggct
     2641 gcaagatgtc ttcctgctcc ctgaccccag tggccagtgg agagatacca gggtctatgg
     2701 cgttttctcc aacccctgga actactcagc tgtctgcgtg tattcgcttg gtgacattga
     2761 cagagtcttc cgtacctcat cgctcaaagg ctaccacatg ggccttccca accctcgacc
     2821 tggcatgtgc ctcccaaaaa agcagcccat acccacagaa accttccagg tagctgatag
     2881 tcacccagag gtggctcaga gggtggaacc tatggggcca ctgaagacac cattgttcca
     2941 ttctaagtac cattaccaga aagtggtcgt tcaccgcatg caagccagca atggagagac
     3001 cttccatgtg ctttatctaa ccacagacag gggcaccatt cacaaggtgg tggaatcagg
     3061 ggaccaggac catagctttg tcttcaatat catggagatc cagccctttc accgtgcagc
     3121 ggccatccag gctatatcat tggatgctga ccggcggaag ctctatgtga cctcccagtg
     3181 ggaagtgagc caggtacccc tggacaaccc taaaccagac gaggctccat tgcagaaggt
     3241 ttccctggct cggaactctc gatactacct gacctgcccc atggagtccc gccacgccac
     3301 ttacttatgg cgccatgagg agaatgtgga acagagctgt gagccaggcc accaaagccc
     3361 tagctgcatc ctgttcattg agaacctcac ggcccgtcag tatggccact accgctgcga
     3421 ggcccaagag ggctcctacc tccgtgaggc tcaacactgg gagctgctgc cagaggacag
     3481 agcactggcc gaacaactaa tgggccatgc ccgggccctg gccgcctcct tctggctggg
     3541 ggtcctgccc acactcatac ttggtctgct ggttcactag gcggccgcta cgtaaattcc
     3601 gccccccccc cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
     3661 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
     3721 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     3781 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     3841 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     3901 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     3961 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     4021 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     4081 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     4141 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg
     4201 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4261 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4321 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4381 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4441 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4501 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4561 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4621 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4681 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4741 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4801 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4861 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     4921 ggccggccgc cagcacagtg gtcgatcgac caatgccctg gctcacaaat accactgaga
     4981 tctttttccc tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc
     5041 tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc
     5101 actcggaagg acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta
     5161 gagtttggca acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg
     5221 tcatcagtat atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac
     5281 ttgaggttag atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa
     5341 attttcctta catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat
     5401 agctgtccct cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat
     5461 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5521 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5581 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct
     5641 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5701 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5761 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5821 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5881 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5941 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     6001 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6061 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6121 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6181 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6241 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6301 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6361 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6421 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6481 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6541 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6601 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6661 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6721 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6781 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6841 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6901 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6961 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7021 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7081 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7141 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7201 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7261 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7321 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7381 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7441 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7501 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7561 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7621 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7681 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7741 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7801 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7861 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7921 taggggttcc gcgcacattt ccccgaaaag tgccacctgg
//
