LOCUS       sequence-304793-        3889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector containing QF2w.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3889)
  AUTHORS   Burgess J, Burrows JT, Sadhak R, Chiang S, Weiss A, D'Amata C, 
            Molinaro AM, Zhu S, Long M, Hu C, Krause HM, Pearson BJ
  TITLE     An optimized QF-binary expression system for use in zebrafish.
  JOURNAL   Dev Biol. 2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007.
  PUBMED    32697972
REFERENCE   2  (bases 1 to 3889)
  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: "Dev Biol. 
            2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007."
COMMENT     Sequence Label: pME-QF2w-SV40pA
FEATURES             Location/Qualifiers
     source          1..3889
                     /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"
     primer_bind     complement(1835..1854)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1889..1908
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1971..1987)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1972..1988)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(2014..2113)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2131..2149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2154..2170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2272..2291)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2283..3092
                     /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(2350..2369)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3239..3827
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3728..3747
                     /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 agcaggctcg aattggagct ccaccgcggg ccgccaccat gccacccaag cgcaaaacgc
      721 ttaacgctgc ggctgaggct aacgctcatg ccgacggaca cgccgacgga aacgccgacg
      781 gacacgtggc caatacggcc gcgtcctcga ataatgcgag gttcgctgat ctcactaaca
      841 tcgatactcc gggtctggga cccacaacta cgaccctgct cgtggaacca gcacgctcaa
      901 agcgtcaacg agtgtcccgc gcatgcgacc agtgccgtgc agcccgagag aaatgcgacg
      961 gaatacagcc tgcgtgtttc ccgtgcgttt cccagggaag gtcctgcact tatcaggctt
     1021 cgccgaaaaa gaggggagtt caaaccggtt atattcgtac gctggagctc gccctcgcct
     1081 ggatgtttga aaatgtcgcg cgttccgaag atgccttgca taacctcctc gtccgtgacg
     1141 ccggacaagg atcagctctg ctcgttggta aagattcgcc ggctgccgag cgactccatg
     1201 cccgttgggc tactagccgt gtcaataaga gcattacccg cctcctccgt cagttggagc
     1261 tccctcctac cgccacggct acggcctcga taatgccgca cgtgatggag cagcctctca
     1321 gtaccagcat taaccccgtc aacgaccgct tcaacggtat tcccaacccc actccgtata
     1381 actccgatgc agctctcgat gctatcactc agaccaacga ttatggaagc gtaaatacac
     1441 atggtatcct ctctacttac ccgccaccgg ctacgcacct taatgaagct tccgtcgctc
     1501 tcgctcccgg tggcgccccc ccccgaccgc ctcctccgta tgttgacagc acgaccaatc
     1561 acccgccgta ccactcgaat ctggttccaa tggcgaactt tggttactcg accgttgatt
     1621 acgatgccat ggttgacgat ttggctagca ttgaatacac ggacgctgtg gatgtcgacc
     1681 cacagtttat gaccaatctg ggattcgttc ctggatgtaa cttctccgac attaatacat
     1741 acaaaaagaa gaaatgaccc gggagcggcc cctctcgagc ctctagaact atagtgagtc
     1801 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1861 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1921 cattataagc tgcaataaac aagagatctg atctagagga tcaagcttat cgataccgtc
     1981 gacctcgagg gggggcccgg tacccagctt ttgacccagc tttcttgtac aaagttggca
     2041 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2101 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2161 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2221 aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     2281 ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg
     2341 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     2401 gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     2461 ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc
     2521 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     2581 ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     2641 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     2701 acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     2761 atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     2821 tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     2881 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     2941 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     3001 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     3061 agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact
     3121 ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg
     3181 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3241 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3301 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3361 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3421 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3481 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3541 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3601 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3661 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3721 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3781 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3841 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
