LOCUS       sequence-99771-e        6206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Middle Entry Vector Containing QS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6206)
  AUTHORS   Subedi A, Macurak M, Gee ST, Monge E, Goll MG, Potter CJ, Parsons 
            MJ, Halpern ME
  TITLE     Adoption of the Q transcriptional regulatory system for zebrafish 
            transgenesis.
  JOURNAL   Methods. 2014 Apr 1;66(3):433-40. doi: 10.1016/j.ymeth.2013.06.012. 
            Epub 2013 Jun 20.
  PUBMED    23792917
REFERENCE   2  (bases 1 to 6206)
  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.ymeth.2013.06.012"; journalName: "Methods"; date: 
            "2014-04-1- 1"; volume: "66"; issue: "3"; pages: "433-40"
COMMENT     Sequence Label: pME-QS (L1-L2)
FEATURES             Location/Qualifiers
     source          1..6206
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     intron          3546..3611
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             3741..3761
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4186..4320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4223..4242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4277..4296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4331..4430)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4447..4466)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4448..4466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4471..4487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4471..4487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4589..4608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4600..5409
                     /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(4667..4686)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5556..6144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6045..6064
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctat gaacaccatc ccggcacgcc atgtcgggga tgtcgccgcc cgcgatcctc
      721 tacctctacc gcacatatca tcctccgtcg ccagcggcat gaagcgttcc ttcgcaacca
      781 tggccatgct ctacaacgac actggcaaca gcaacgatgt cggtgcccat gccaggcgac
      841 caccacgaac cctctccaat agtcggagca cttccgccca cagagtacct ctaggctctt
      901 ggtcggcgcc caattccccg ccccgccgtg cgctgccgca tcatcccatc accgccagct
      961 tcgatcccga tgcttccatt gttatcgccg gcattcgtgg cgctggcaag tctacgctgg
     1021 ccatcatggc atctaccgcc atgaagcgca agatagtcga cctggaatcc gagttccatc
     1081 atcttactgg cttgtctagt tccagctaca agaagacaca cggcccggtc gactatggga
     1141 ggcgccagat cgccatcttg cagaacatct tgaatctgca caggacccgc gccattctcg
     1201 tctgctcttg gctggagcgg gatgtgcagg ccatgttgca ggatttcagc gtgtctaatc
     1261 ctgtcattta cgttctgcgc gatgccaaag ctatcgaggc ccatctgaag ggatacgaca
     1321 agtccaaagt tggcaccctc cttgatgcca ccagtactgt ccttcgccgc tgcacccgtt
     1381 tcgagttctt caacgtctcc gaagagaacc tggacaccca ctccgcttca acatcaccac
     1441 ctgctgttcc ggaccagcgg cataccgcgc cgtatctaac gcttaaacga gccgagcgcc
     1501 acttcctcaa atttctctcc ttgattctac ccaaggggac catacctttt gtcgagtccg
     1561 cctttcccct ggcttccgtc cccgttgaac agcgccgctt tacctacgcc ctcgccttgc
     1621 ctgtatctgc cttgctcgac aaaggcgtcg atatccaaga gcttgatgtc ggtgtagacg
     1681 caatcgagat cattgtagac gatcttgcaa cgagcgaatc cggcccaacg agccccttgg
     1741 gtcttgcgcc ccaccgagcg agcgagatca gtcgtgttgt aggcgaaatc aggagggaca
     1801 cagtgatccc catcattctg cacgtggtct ttccagaaag agcgctatat gaagaagctc
     1861 tgctcgcgct ttacatgact tacctgaacc atgccttaag gcttgcgcca gattatctta
     1921 cggtcgatct ggggctcgat tctggcttgc ttgggcaact aaccaccgtt caaggaacca
     1981 ccaaggtcat cggcaataaa caacttgcag aggtcaattc gccgcgctgg ggggatccat
     2041 cttggttaca agcctatgaa aaggcccaga atacaggatg cgacttggta aggttgacca
     2101 gaccggcttc aaatccccgg gacaacacag acattcggca gttccacgtt gctgtagagg
     2161 ccgtcggggg tccaaggctc ccatttattg cttacaacac aggacgccta ggtcggacat
     2221 cgatgtgttt taacgagatc ctgactccag ttacaccagt gcctaccaag gaggatgcaa
     2281 tcgggctccg caatccagcc catcgctatc tccagcctcc gctcacggct ctggaagcaa
     2341 cacaggctct ctactcggca tttgtccacg acccaatgaa gctgtatgtc tttggcgcaa
     2401 atgtgggata tagcttgtcc ccagccatgc acaatgccgc actcaaggcc tgtggcattc
     2461 cacaccatta caagcccctt tccacagcaa acatcgggac tttgcgcgag gttatcagcg
     2521 atccgcagtt tgctggagcc tcggtcggcc tgccgtttaa ggtggaaatc atcagcctca
     2581 cacactcgct gagccggcac gcgaaagcca tcggagccgt caacaccttg attccggtac
     2641 gacaccttac cgcggacggt ggaataccgg acgaggtgtc catgttcaac aatatcagcc
     2701 aagccggcgc tgtcagagct ctctacggcg agaacacgga ttggattggt atccgagcct
     2761 gccttcgccg cggtttatcg cccgccaatg ccgtgagatc aacaagcact ggtcttgtca
     2821 tcggcgctgg cggaatggct agggcagctg tctatgccat gcttcaactg ggagtcaaga
     2881 agattttgat ctttaaccga acatttgcta atgccgagaa gctggttcta cacttcgaga
     2941 acctgttggt cagagacgca ttgcctctgt tgagcacagg gccaagatcc cacgacaaca
     3001 cctgttttca catcattcga tctcgagacg atccgctccc agaaaacttc aaaaacccga
     3061 ccatgatcgt ttcctgcata ccgacacaca cagtggacaa cacccctgac cctgaattta
     3121 ctgtgccttt gcactggctc gacaacccca ctggcggcat tgtactagaa ctcgactaca
     3181 aatgtctcac atcacccttg ctcgaacaaa cacgacgcga ggctcacaga ggctgggtcg
     3241 caatggatgg acttgacctc ttgccagaac aagggtttgc ccaatttgaa ctgttcaccg
     3301 ggcggcgagc acctcgtcgc ttgatgaggc gcgaggtttt gcgagcatac ccagatgatc
     3361 aagcaaaatc tcataccgcg cagttacagc ctcgcctcaa cggaattgca acgcaaatat
     3421 cttgagccta gatccgcgac ccatttgctg tccaccagtc atgctagagg atctttgtga
     3481 aggaacctta cttctgtggt gtgacataat tggacaaact acctacagag atttaaagct
     3541 ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     3601 gtgtatttta gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta
     3661 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg
     3721 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc
     3781 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct
     3841 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat
     3901 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta
     3961 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta
     4021 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag
     4081 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4141 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4201 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4261 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4321 ggatccaggc acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     4381 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     4441 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     4501 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata
     4561 aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa
     4621 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     4681 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     4741 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     4801 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     4861 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta
     4921 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     4981 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     5041 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     5101 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     5161 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     5221 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     5281 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     5341 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     5401 tttttctaat cagaattggt taattggttg taacactggc agagcattac gctgacttga
     5461 cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga
     5521 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5581 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5641 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5701 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5761 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5821 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5881 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5941 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6001 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6061 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6121 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6181 ttttgctggc cttttgctca catgtt
//
