LOCUS       sequence-341942-        4468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector for Gateway cloning that contains a ReaChR fused
            with citrine gene insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4468)
  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 4468)
  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"; 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: pME-ReaChR-citrine
FEATURES             Location/Qualifiers
     source          1..4468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(4..103)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(120..139)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(121..139)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(144..160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(144..160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(262..281)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             273..1082
                     /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(340..359)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1229..1817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1718..1737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1971..1988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2147..2174)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2266..2352)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2401..2423
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2415..2432
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2416..2432
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2448..2547
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(3752..3754,3755..3757,3758..4468)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3797..3818)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4058..4077)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4405..4426
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ccgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt
       61 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc
      121 cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc
      181 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
      241 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga
      301 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
      361 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt
      421 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
      481 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
      541 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag
      601 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
      661 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
      721 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
      781 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
      841 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
      901 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
      961 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     1021 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     1081 aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg
     1141 gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     1201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     1921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     1981 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2041 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2101 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     2161 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     2221 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     2281 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     2341 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt
     2401 cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt
     2461 tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa
     2521 tgccaacttt gtacaaaaaa gcaggctccg cggccaatct gctcagagag gacagagtgg
     2581 gcaggagcca gcattgggta tataaagctg agcagggtca gttgcttctt acgtttgctt
     2641 ctgagtctgt tgtgttgact tgcaacctca gaaacagaca tccgccccct tcaccatggt
     2701 gagcagaaga ccctggctgc tggccctggc cctggccgtg gccctggccg ccggcagcgc
     2761 cggcgccagc accggcagcg acgccaccgt gcccgtggcc acccaggacg gccccgacta
     2821 cgtgttccac agagcccacg agagaatgct gttccagacc agctacaccc tggagaacaa
     2881 cggcagcgtg atctgcatcc ccaacaacgg ccagtgcttc tgcctggcct ggctgaagag
     2941 taacggcacc aacgccgaga agctggccgc caacatcctg cagtgggtgg tgtttgcgct
     3001 gagcgtggcg tgcctgggct ggtatgcgta tcaggcgtgg cgcgcgacct gcggctggga
     3061 agaagtgtat gtggcgctga ttgaaatgat gaaaagcatt attgaagcgt ttcatgaatt
     3121 tgatagcccg gcgaccctgt ggctgagcag cggcaacggc gtggtgtgga tgcgctatgg
     3181 cgaatggctg ctgacctgcc ccgtgattct gattcatctg agcaacctga ccggcctgaa
     3241 agatgattat agcaaacgca ccatgggcct gctggtgagc gacgtgggct gcattgtgtg
     3301 gggcgcgacc agcgcgatgt gcaccggctg gaccaaaatt ctgttttttc tgattagcct
     3361 gagctatggc atgtatacct attttcatgc ggccaaagtg tatattgaag cgtttcatac
     3421 cgtgccgaaa ggcctgtgca gacagctggt gagagccatg gcctggctgt tcttcgtgag
     3481 ctggggcatg ttccccgtgc tgttcctgct gggccccgag ggcttcggcc atattagccc
     3541 gtatggcagc gcgattggcc atagcattct ggatctgatt gcgaagaaca tgtggggcgt
     3601 gctgggcaac tatctgcgcg tgaaaattca tgaacatatt ctgctgtatg gcgatattcg
     3661 caaaaaacag aaaattacca ttgcgggcca ggaaatggaa gtggaaaccc tggtggcgga
     3721 agaagaagat aagtacgaga gcagcctcga gatggtgagc aagggcgagg agctgttcac
     3781 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     3841 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     3901 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgat
     3961 gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4021 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4081 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4141 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4201 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4261 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4321 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa
     4381 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     4441 cactctcggc atggacgagc tgtacaag
//
