LOCUS       sequence-310847-        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"
     terminator      complement(92..178)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     227..249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     241..258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     242..258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    274..373
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1540..1559)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1594..1613
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1676..1692)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1677..1693)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(1719..1818)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1836..1854)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1859..1875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1859..1875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1977..1996)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1988..2797
                     /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(2055..2074)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2944..3532
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3433..3452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3686..3703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3862..3889)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
       61 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
      121 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
      181 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
      241 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
      301 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac
      361 aaaaaagcag gctcgaattg gagctccacc gcgggccgcc accatgccac ccaagcgcaa
      421 aacgcttaac gctgcggctg aggctaacgc tcatgccgac ggacacgccg acggaaacgc
      481 cgacggacac gtggccaata cggccgcgtc ctcgaataat gcgaggttcg ctgatctcac
      541 taacatcgat actccgggtc tgggacccac aactacgacc ctgctcgtgg aaccagcacg
      601 ctcaaagcgt caacgagtgt cccgcgcatg cgaccagtgc cgtgcagccc gagagaaatg
      661 cgacggaata cagcctgcgt gtttcccgtg cgtttcccag ggaaggtcct gcacttatca
      721 ggcttcgccg aaaaagaggg gagttcaaac cggttatatt cgtacgctgg agctcgccct
      781 cgcctggatg tttgaaaatg tcgcgcgttc cgaagatgcc ttgcataacc tcctcgtccg
      841 tgacgccgga caaggatcag ctctgctcgt tggtaaagat tcgccggctg ccgagcgact
      901 ccatgcccgt tgggctacta gccgtgtcaa taagagcatt acccgcctcc tccgtcagtt
      961 ggagctccct cctaccgcca cggctacggc ctcgataatg ccgcacgtga tggagcagcc
     1021 tctcagtacc agcattaacc ccgtcaacga ccgcttcaac ggtattccca accccactcc
     1081 gtataactcc gatgcagctc tcgatgctat cactcagacc aacgattatg gaagcgtaaa
     1141 tacacatggt atcctctcta cttacccgcc accggctacg caccttaatg aagcttccgt
     1201 cgctctcgct cccggtggcg cccccccccg accgcctcct ccgtatgttg acagcacgac
     1261 caatcacccg ccgtaccact cgaatctggt tccaatggcg aactttggtt actcgaccgt
     1321 tgattacgat gccatggttg acgatttggc tagcattgaa tacacggacg ctgtggatgt
     1381 cgacccacag tttatgacca atctgggatt cgttcctgga tgtaacttct ccgacattaa
     1441 tacatacaaa aagaagaaat gacccgggag cggcccctct cgagcctcta gaactatagt
     1501 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     1561 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1621 gtaaccatta taagctgcaa taaacaagag atctgatcta gaggatcaag cttatcgata
     1681 ccgtcgacct cgaggggggg cccggtaccc agcttttgac ccagctttct tgtacaaagt
     1741 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca
     1801 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt
     1861 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1921 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1981 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     2041 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     2101 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     2161 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     2221 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     2281 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     2341 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     2401 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2461 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2521 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2581 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2641 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2701 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2761 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2821 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2881 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2941 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     3001 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     3061 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     3121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3541 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3721 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3781 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     3841 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttct
//
