LOCUS       sequence-215007-        4389 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 4389)
  TITLE     p3E_he1a constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4389)
  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            "Unpublished"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..4389
                     /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    complement(624..748)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1188..1898
                     /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(1333..1351)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1601..1620
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1795..1818
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2265..2282)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2271..2496
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(2498..2593)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2630..2649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2631..2649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2654..2670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2654..2670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2772..2791)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2783..3592
                     /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(2850..2869)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3739..4327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4228..4247
                     /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggtca accactccag gcatagctag agccactgat
      781 gcatctatag agctgaagta aagatagggt tgatgcaaat cgatttagaa ttttcaatgt
      841 agaaaaaaca aaataaaaag tctattcact tgatggatag attagtcaaa caaacataga
      901 gtaggcactt cccttgactc tgtaaacttt tggatttaac agattgtgca ctccttttag
      961 ttaaacacat tagtccctga aaactaagac cctaaccata atcccattgt agacccacct
     1021 actcgggtga atataaagct cgcacagatc atctcacaga catcagacct tggcaaagga
     1081 gcatcctgaa catctctaca cactgaggca accctagggc cgccaccatg ctgtgttgta
     1141 tcagaagaac aaagcctgtg gagaagaatg aggaggctga ccaggaaatg gtgagcaagg
     1201 gcgaggagga caacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     1261 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     1321 gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca
     1381 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     1441 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     1501 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     1561 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     1621 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     1681 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgccgag gtcaagacca
     1741 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagctgg
     1801 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc gccgagggcc
     1861 gccactccac cggcggcatg gacgagcttt acttgtaagt aaagcggccg cgactctaga
     1921 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1981 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttatcgaa ttctgcagga
     2041 attgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact
     2101 ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg
     2161 tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag
     2221 acaacctgta gggcacgagg gggggcccga aacccgctga tcagcctcga ctgtgccttc
     2281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     2461 tagcaaggca tgctggggat gcggtgggct ctatggccaa ctttattata caaagttggc
     2521 attataaaaa agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat
     2581 aaaatcatta tttggagctc catggtagcg ttaacgcggc cgcgatatcc cctatagtga
     2641 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2701 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     2761 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     2821 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     2881 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     2941 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     3001 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     3061 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     3121 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     3181 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     3241 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     3301 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     3361 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     3421 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     3481 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     3541 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     3601 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     3661 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     3721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3841 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     4021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag
     4141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4381 tcacatgtt
//
