LOCUS       sequence-484424-        3091 bp    DNA     circular SYN 22-AUG-2025
DEFINITION  5' entry vector for Gateway cloning for expression in spinal motor 
            neurons using 3 tandem repeats of the zebrafish mnx1 enhancer..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3091)
  AUTHORS   Harris JM, Wang AY, Boulanger-Weill J, Santoriello C, Foianini S, 
            Lichtman JW, Zon LI, Arlotta P
  TITLE     Long-Range Optogenetic Control of Axon Guidance Overcomes 
            Developmental Boundaries and Defects.
  JOURNAL   Dev Cell. 2020 Jun 8;53(5):577-588.e7. doi: 
            10.1016/j.devcel.2020.05.009.
  PUBMED    32516597
REFERENCE   2  (bases 1 to 3091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 22, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2020.05"; journalName: "Dev Cell"; date: 
            "2020-06-8- 8"; volume: "53"; issue: "5"; pages: "577-588.e7"
COMMENT     Sequence Label: p5E-3x125bp-mnx1
FEATURES             Location/Qualifiers
     source          1..3091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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"
     protein_bind    1133..1256
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1332..1351)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1333..1351)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1356..1372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1356..1372)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1474..1493)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1485..2294
                     /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"
     primer_bind     complement(1552..1571)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2441..3029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2930..2949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 tcgggcccga gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggc tccgaattcg ccctttcctc gagagtggtt
      721 agctgatgaa ttgaggaaaa ctaatccgct ttattggtag acaggttaag ggcaacagag
      781 tgtcaataat tctctccctg acctctttct ccattagctt aaagaggctt ataagagggc
      841 atgccgagag tggttagctg atgaattgag gaaaactaat ccgctttatt ggtagacagg
      901 ttaagggcaa cagagtgtca ataattctct ccctgacctc tttctccatt agcttaaaga
      961 ggcttataag aggcctaggc gagagtggtt agctgatgaa ttgaggaaaa ctaatccgct
     1021 ttattggtag acaggttaag ggcaacagag tgtcaataat tctctccctg acctctttct
     1081 ccattagctt aaagaggctt ataagaggaa gcttgtaagg gcgaattcga cccaagtttg
     1141 tacaaaaaag ttgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt
     1201 ttgcataaaa aacagactac ataatactgt aaaacacaac atatgcagtc actatgaacc
     1261 aactacttag atggtattag tgacctgtag aattaattcg agctctagag ctgcagggcg
     1321 gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1381 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
     1441 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
     1501 gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     1561 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg
     1621 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     1681 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     1741 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc
     1801 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     1861 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     1921 gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     1981 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat
     2041 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     2101 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     2161 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     2221 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     2281 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     2341 cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     2401 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2461 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2521 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2581 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2641 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2701 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2761 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2821 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2881 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2941 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3001 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3061 tggccttttg ctggcctttt gctcacatgt t
//
