LOCUS       sequence-38084-e        4523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4523)
  AUTHORS   White MD, Milne RV, Nolan MF
  TITLE     A Molecular Toolbox for Rapid Generation of Viral Vectors to Up- or 
            Down-Regulate Neuronal Gene Expression in vivo.
  JOURNAL   Front Mol Neurosci. 2011;4:8. Epub 2011 Jul 4.
  PUBMED    21772812
REFERENCE   2  (bases 1 to 4523)
  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"; journalName: "Front Mol 
            Neurosci."; date: "2011"; volume: "4"; pages: "8. Epub 2011 Jul 4"
COMMENT     Sequence Label: pENTR-L5-Kir2.1-mCherry-L2
FEATURES             Location/Qualifiers
     source          1..4523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             343..354
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    385..480
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      480..489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1779..2489
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1924..1942)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2192..2211
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2500..2599)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2729..2746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2839..3427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2919..2938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3601..4395)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4396..4500)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattgaagg aaggccgtca
      361 aggcctaggc gcgcctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      421 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtat acaaaagttg
      481 ccgccatggg cagtgtgaga accaaccgct acagcatcgt ctcttcggag gaagatggca
      541 tgaagctggc cactatggca gttgccaatg gctttgggaa tggcaagagt aaagtccata
      601 cccgacaaca gtgcaggagc cgctttgtga agaaagatgg tcattgcaat gttcagttta
      661 tcaacgtggg tgagaaggga cagaggtacc tggcagacat ctttactacc tgtgtcgaca
      721 tccgctggag gtggatgctg gttatcttct gtcttgcctt cgtgctctcc tggctgttct
      781 ttggctgtgt gttttggttg atagccctgc tccatgggga tctagatact tctaaagtga
      841 gcaaagcatg cgtgtcagag gtcaacagct tcacggctgc cttcctcttc tccatcgaga
      901 cccagacaac cattggctat ggtttcaggt gtgtgacaga cgagtgccca attgctgtct
      961 tcatggtggt attccagtca atcgtaggct gcatcattga cgccttcatc attggtgcag
     1021 tcatggcgaa gatggcaaag ccaaagaaga gaaatgagac tcttgtcttc agtcacaatg
     1081 ctgtgattgc catgagggat ggcaaactct gcttgatgtg gagagtgggt aaccttcgaa
     1141 agagccacct tgtggaagct catgtccggg cacagcttct caaatctagg atcacttcag
     1201 aaggggagta tatccctttg gaccagatag acatcaatgt tggttttgat agtggaattg
     1261 accgcatatt tctagtgtcc cccatcacta tcgttcacga aatagatgaa gacagccctt
     1321 tatatgactt gagtaagcag gacattgaca atgcagactt tgaaattgtt gtcatactgg
     1381 aaggcatggt ggaggcgact gccatgacaa ctcaatgccg gagttcgtat ctggccaatg
     1441 aaattctctg gggtcaccgc tatgagccag tgctctttga agagaaacac tactataaag
     1501 tagactattc aagattccat aagacttatg aagtacctaa cacccccctt tgtagtgcca
     1561 gagacttagc agagaagaaa tacatccttt caaatgcaaa ttcattttgc tatgaaaatg
     1621 aagttgccct aacaagcaaa gaggaagagg aggatagtga gaacggagtc ccagagagca
     1681 caagcacaga ctcacctcct ggcatagatc tccacaacca ggcaagcgta cctctagagc
     1741 ccaggccctt aaggcgagaa tcggagatag ggccggccat ggtgagcaag ggcgaggagg
     1801 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1861 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1921 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1981 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     2041 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     2101 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     2161 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     2221 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     2281 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     2341 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     2401 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     2461 ccggcggcat ggacgagctg tacaagtgag acgctgtcta cccagctttc ttgtacaaag
     2521 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     2581 aaaataaaat cattatttgc catccagtta attaactggc ctcatgggcc ttcctttcac
     2641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt
     2701 tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2761 gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3601 ttagaaaaat tcatccagca gacgataaaa cgcaatacgc tggctatccg gtgccgcaat
     3661 gccatacagc accagaaaac gatccgccca ttcgccgccc agttcttccg caatatcacg
     3721 ggtggccagc gcaatatcct gataacgatc cgccacgccc agacggccgc aatcaataaa
     3781 gccgctaaaa cggccatttt ccaccataat gttcggcagg cacgcatcac catgggtcac
     3841 caccagatct tcgccatccg gcatgctcgc tttcagacgc gcaaacagct ctgccggtgc
     3901 caggccctga tgttcttcat ccagatcatc ctgatccacc aggcccgctt ccatacgggt
     3961 acgcgcacgt tcaatacgat gtttcgcctg atgatcaaac ggacaggtcg ccgggtccag
     4021 ggtatgcaga cgacgcatgg catccgccat aatgctcact ttttctgccg gcgccagatg
     4081 gctagacagc agatcctgac ccggcacttc gcccagcagc agccaatcac ggcccgcttc
     4141 ggtcaccaca tccagcaccg ccgcacacgg aacaccggtg gtggccagcc agctcagacg
     4201 cgccgcttca tcctgcagct cgttcagcgc accgctcaga tcggttttca caaacagcac
     4261 cggacgaccc tgcgcgctca gacgaaacac cgccgcatca gagcagccaa tggtctgctg
     4321 cgcccaatca tagccaaaca gacgttccac ccacgctgcc gggctacccg catgcaggcc
     4381 atcctgttca atcatactct tcctttttca atattattga agcatttatc agggttattg
     4441 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4501 cacatttccc cgaaaagtgc cac
//
