LOCUS       sequence-412271-        6523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SLIT2 protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6523)
  TITLE     TREAT-AD plasmids Jun 2023
REFERENCE   2  (bases 1 to 6523)
  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"
COMMENT     Sequence Label: SLIT2A-c003
FEATURES             Location/Qualifiers
     source          1..6523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          640..1656
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1580..1602
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1664..1683
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1738..1747
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3688..3732
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3745..3762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3874..3893)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3939..3994
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3993..4012)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4355..4371)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4355..4371)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4368..4390)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4379..4395
                     /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(4403..4409,4410..4427,4428..4433))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4448..4469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4598..4615)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4769..5357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4849..4868)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5528..6319,6320..6388))
                     /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     6151..6170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6389..6493)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggggcgcgc gggggggggg gsggggcgrg gcggagaggt
      541 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      601 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      661 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      721 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      781 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      841 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
      901 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      961 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
     1021 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     1081 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
     1141 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
     1201 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
     1261 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
     1321 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     1381 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1441 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1501 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1561 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct
     1621 gctaaccatg ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt
     1681 gtgctgtctc atcattttgg caaagaattg cggccgtctc aggccgaatt caagcttgcc
     1741 accatgggga tccttcccag ccctgggatg cctgcgctgc tctccctcgt gagccttctc
     1801 tccgtgctgc tgatgggttg cgtagctgaa accggtccct gcctatcaaa tccgtgtaaa
     1861 aatgatggca catgtaatag tgatccagtt gacttttacc gatgcacctg tccatatggt
     1921 ttcaaggggc aggactgtga tgtcccaatt catgcctgca tcagtaaccc atgtaaacat
     1981 ggaggaactt gccacttaaa ggaaggagaa gaagatggat tctggtgtat ttgtgctgat
     2041 ggatttgaag gagaaaattg tgaagtcaac gttgatgatt gtgaagataa tgactgtgaa
     2101 aataattcta catgtgtcga tggcattaat aactacacat gcctttgccc acctgagtat
     2161 acaggtgagt tgtgtgagga gaagctggac ttctgtgccc aggacctgaa cccctgccag
     2221 cacgattcaa agtgcatcct aactccaaag ggattcaaat gtgactgcac accagggtac
     2281 gtaggtgaac actgcgacat cgattttgac gactgccaag acaacaagtg taaaaacgga
     2341 gcccactgca cagatgcagt gaacggctat acgtgcatat gccccgaagg ttacagtggc
     2401 ttgttctgtg agttttctcc acccatggtc ctccctcgta ccagcccctg tgataatttt
     2461 gattgtcaga atggagctca gtgtatcgtc agaataaatg agccaatatg tcagtgtttg
     2521 cctggctatc agggagaaaa gtgtgaaaaa ttggttagtg tgaattttat aaacaaagag
     2581 tcttatcttc agattccttc agccaaggtt cggcctcaga cgaacataac acttcagatt
     2641 gccacagatg aagacagcgg aatcctcctg tataagggtg acaaagacca tatcgcggta
     2701 gaactctatc gggggcgtgt tcgtgccagc tatgacaccg gctctcatcc agcttctgcc
     2761 atttacagtg tggagacaat caatgatgga aacttccaca ttgtggaact acttgccttg
     2821 gatcagagtc tctctttgtc cgtggatggt gggaacccca aaatcatcac taacttgtca
     2881 aagcagtcca ctctgaattt tgactctcca ctctatgtag gaggcatgcc agggaagagt
     2941 aacgtggcat ctctgcgcca ggcccctggg cagaacggaa ccagcttcca cggctgcatc
     3001 cggaaccttt acatcaacag tgagctgcag gacttccaga aggtgccgat gcaaacaggc
     3061 attttgcctg gctgtgagcc atgccacaag aaggtgtgtg cccatggcac atgccagccc
     3121 agcagccagg caggcttcac ctgcgagtgc caggaaggat ggatggggcc cctctgtgac
     3181 caacggacca atgacccttg ccttggaaat aaatgcgtac atggcacctg cttgcccatc
     3241 aatgcgttct cctacagctg taagtgcttg gagggccatg gaggtgtcct ctgtgatgaa
     3301 gaggaggatc tgtttaaccc atgccaggcg atcaagtgca agcatgggaa gtgcaggctt
     3361 tcaggtctgg ggcagcccta ctgtgaatgc agcagtggat acacggggga cagctgtgat
     3421 cgagaaatct cttgtcgagg ggaaaggata agagattatt accaaaagca gcagggctat
     3481 gctgcttgcc aaacaaccaa gaaggtgtcc cgattagagt gcagaggtgg gtgtgcagga
     3541 gggcagtgct gtggaccgct gaggagcaag cggcggaaat actctttcga atgcactgac
     3601 ggctcctcct ttgtggacga ggttgagaaa gtggtgaagt gcggctgtac gaggtgtgtg
     3661 tccggtaccg gaggttccgg tggttccggt ctgaatgata tctttgaagc tcagaagatt
     3721 gaatggcatg aaggacgtac caagcaccac catcaccatc actaatgatc actcgagact
     3781 agtatcgcga taattcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3841 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3901 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3961 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     4021 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     4081 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     4141 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     4201 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     4261 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     4321 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4381 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4441 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4501 cgggaaacct gtcgtgccag cggatcgatc cgctgcatta atgaatcggc caacgcgcgg
     4561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6361 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6421 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6481 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtc
//
