LOCUS       Exported                4747 bp ds-DNA     circular SYN 23-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 4747)
  AUTHORS   Wythe Lab
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 22, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4747
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     terminator      268..295
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     667..718
                     
     enhancer        717..2785
                     /label= hs GLAST promotor
                     /note="GenBank: AF448436.1"
     primer_bind     740..762
                     /label=oJDW 755
     primer_bind     complement(740..762)
                     /label=oJDW 756
     primer_bind     complement(1323..1355)
                     /label=oJDW 1721
     primer_bind     1389..1411
                     /label=oJDW 757
     primer_bind     complement(1902..1922)
                     /label=oJDW 761
     primer_bind     1964..1983
                     /label=oJDW 758
     primer_bind     2475..2494
                     /label=oJDW 852 (GLAST FWD)
     primer_bind     2722..2741
                     /label=oJDW 759
     primer_bind     complement(2722..2741)
                     /label=oJDW 760
     primer_bind     complement(2766..2810)
                     /label=oJDW 2071 (attB1- hsGLAST-REV-AgeI-SpeI)
     protein_bind    2793..2916
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3012..3028)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3141..3950
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4097..4685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgta tcgataggta ccatgtctac acaaactgaa
      721 ataatgaaat ggtttttaac caagctttag ggattagaga ggaaaaaaaa agtgaaaaag
      781 aaagggcacc agggtcttcc catctctatg ctaaccaaag tgtggttttc cagccctttt
      841 tcttgcctat taacaaaaat aaaaacaaaa aatggaaatg ccactgggct tgaaacccca
      901 tttaaaatca gatttctctt tcgagttccc ttgcaataca agtcaaagtc atttccacca
      961 aaaaaaaaaa gcggtggggg ggggtgcgca aattaatcct aaactcatgg ctcagatagg
     1021 acctggtata gtaaataaat gcttttaaaa gtagtcatgg cattgagaaa tggctccatg
     1081 cacctaaacc agaggcacca atgtaaaaac tagtaattgt gttttaacac aaaagattga
     1141 aaagcgtttt gtttcacatc ctaatattat tccacccact taaaaagagt tacttaatat
     1201 tgactctgta ttaggtataa gacaagttct tgccctcaaa gaacttataa tttagttgag
     1261 aaaccaaaac ttaacatgag cttaaaaaga aggatgcagt acaaaacatg gtataattaa
     1321 gtgctaaaca atgtagtata gatgctcagt gacctaggag atcagaggaa gggaagggtc
     1381 actgagggct ggggatgtca aaagcatgta ctttcagtaa gttcttaaaa tgatagcatg
     1441 ttagaaagct gttataattg ctgctgattt gtaattttaa agcgttgaca tgattcacca
     1501 ttgctttcca accagtagct gtttaaaagg acatacagtt gagggtgaaa ggagatattt
     1561 ctttatgatc tccagagatt tgtattgagg ggaaaaaaga catactttct tgttgaataa
     1621 tagaccaaga attgagcata ttttgcatgg ggaatattaa attcagtgaa caagaatcca
     1681 aacctcaggc tgtgattctg gtataaattt ttgtttctaa tgaaccattt tcttccttct
     1741 aaagaagatt taagttttgg gaataagtca cctcaaaaaa tttttttttt attttctaat
     1801 atctattttc acaatttatt tttdttttaa gaaaagtgga ctagaagctc aagttacctt
     1861 ctcaaggcaa tgagaatttt ttagctctaa aaaagttgtg tgtggtaggg agaacattta
     1921 gcatccttta aaatggagac atctgaacag caacctatct tttgaatcgc caaatctcaa
     1981 gtgtactttg cactctcggt cttaaaatgc tcttcatttt ctagtttggc aaaaattcca
     2041 gtctctttac atttgctacc ttttccgtct ttgcaacaca ggcagtgtga gctaaattag
     2101 gtttgttaaa atgaagtttt gaagtttcaa ctgctgaaat agaggcatgt ccctaacttt
     2161 agacaaattt agagaatcgt agtaagcatt attttttttc tacatacttg tctgcattca
     2221 gtgtcttccc aagaaacgtc agttaattcc ccgattcgaa aaaggaactt tctcgtaaca
     2281 gttgtacaac caggctaaag agcacaccct cgtcttccct gaaagggagg aaacatgcca
     2341 taatgtgagg taattaaatt tagccttaga aaagtctttg aaacgttcta atcaagactt
     2401 tcagctttcc gattgtgtgc gctttatttt tcaattcttc ctagtcccgg ttttgcattt
     2461 ctaatgacag tgaactgaca aagcgtgaaa gtggtctaag gagcaaaaca aagccagcaa
     2521 gcctgcctct ggtgtgccaa gattaatcaa tattccagaa acctcggggt ttccccctcc
     2581 tccctgtgcg gtttccattc cccactcctc ctttcccggg cacagactcc tccccttttc
     2641 ttaggttgcc ctgatagtaa cttgcggttt cagagcacat gcacactgtc agggctagcc
     2701 tgcctgctta cgcgcgctgc ggattgctgc tccgttgtac ctgctgggga attcacctcg
     2761 ttactgcttg atgggctcga gatccaccgg ttcaagtttg tacaaaaaag ttgaacgaga
     2821 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     2881 ataatactgt aaaacacaac atatgcagtc actatgaatc aactacttag atggtattag
     2941 tgacctgtag aattcgagct ctagagctgc agggcggccg cgatatcccc tatagtgagt
     3001 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     3061 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     3121 acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa
     3181 attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat
     3241 caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac
     3301 atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga
     3361 cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt
     3421 tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt
     3481 caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg
     3541 tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa
     3601 tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg
     3661 aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc
     3721 atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg
     3781 atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc
     3841 tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc
     3901 ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg
     3961 ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc gcaagctcat
     4021 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
     4081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     4261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     4501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4681 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4741 acatgtt
//
