LOCUS       sequence-223633-        4388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  p3E entry vector for Three Multisite Gateway.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4388)
  TITLE     p3E_he1a constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4388)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p3E_he1a:mCherry
FEATURES             Location/Qualifiers
     source          1..4388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(23..47)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             487..1197
                     /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
                     EGRHSTGGMDELYL"
     primer_bind     complement(632..650)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     900..919
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1094..1117
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1564..1581)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1570..1795
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1796..1891)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1928..1947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1929..1947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1952..1968)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1952..1968)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2070..2089)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2081..2890
                     /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(2148..2167)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3037..3625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3526..3545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3779..3796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3955..3982)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4074..4160)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4209..4231
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4223..4240
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4224..4240
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tttatatcat tttacgtttc tcgttcaact ttcttgtaca aagtggtcaa ccactccagg
       61 catagctaga gccactgatg catctataga gctgaagtaa agatagggtt gatgcaaatc
      121 gatttagaat tttcaatgta gaaaaaacaa aataaaaagt ctattcactt gatggataga
      181 ttagtcaaac aaacatagag taggcacttc ccttgactct gtaaactttt ggatttaaca
      241 gattgtgcac tccttttagt taaacacatt agtccctgaa aactaagacc ctaaccataa
      301 tcccattgta gacccaccta ctcgggtgaa tataaagctc gcacagatca tctcacagac
      361 atcagacctt ggcaaaggag catcctgaac atctctacac actgaggcaa ccctagggcc
      421 gccaccatgc tgtgttgtat cagaagaaca aagcctgtgg agaagaatga ggaggctgac
      481 caggaaatgg tgagcaaggg cgaggaggac aacatggcca tcatcaagga gttcatgcgc
      541 ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc
      601 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      661 ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg
      721 aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg
      781 gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      841 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
      901 cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
      961 gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac
     1021 gacgccgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1081 aacgtcaaca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1141 tacgagcgcg ccgagggccg ccactccacc ggcggcatgg acgagcttta cttgtaagta
     1201 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     1261 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1321 gttatcgaat tctgcaggaa ttgggtggca tccctgtgac ccctccccag tgcctctcct
     1381 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     1441 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     1501 caaggggcaa gttgggaaga caacctgtag ggcacgaggg ggggcccgaa acccgctgat
     1561 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1621 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1681 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1741 gggaggattg ggaagacaat agcaaggcat gctggggatg cggtgggctc tatggcaact
     1801 ttattataca aagttggcat tataaaaaag cattgcttat caatttgttg caacgaacag
     1861 gtcactatca gtcaaaataa aatcattatt tggagctcca tggtagcgtt aacgcggccg
     1921 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1981 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     2041 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     2101 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2161 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2221 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2281 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2341 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2401 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2461 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2521 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2581 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2641 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2701 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2761 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2821 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2881 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2941 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     3001 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3061 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3661 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3721 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3781 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3841 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3901 gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg
     3961 ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct
     4021 gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc
     4081 ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac
     4141 tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg
     4201 gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccctgcag
     4261 ctctagagct cgaattctac aggtcactaa taccatctaa gtagttggtt catagtgact
     4321 gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata
     4381 tattgata
//
