LOCUS       sequence-222563-        4313 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 4313)
  TITLE     p3E_he1a constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4313)
  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            "Unpublished"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:eYFP
FEATURES             Location/Qualifiers
     source          1..4313
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(52..176)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(661..663,664..666,667..1380)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(706..727)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(967..986)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1314..1335
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1618..1635)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1624..1849
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(1850..1945)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1982..2001)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1983..2001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2006..2022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2006..2022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2124..2143)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2135..2944
                     /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(2202..2221)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3091..3679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3580..3599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3833..3850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4009..4036)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4128..4214)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4263..4285
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4277..4294
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4278..4294
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gctctagagc tcgaattcta caggtcacta ataccatcta agtagttggt tcatagtgac
       61 tgcatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
      121 atattgatat ttatatcatt ttacgtttct cgttcaactt tcttgtacaa agtggtcaac
      181 cactccaggc atagctagag ccactgatgc atctatagag ctgaagtaaa gatagggttg
      241 atgcaaatcg atttagaatt ttcaatgtag aaaaaacaaa ataaaaagtc tattcacttg
      301 atggatagat tagtcaaaca aacatagagt aggcacttcc cttgactctg taaacttttg
      361 gatttaacag attgtgcact ccttttagtt aaacacatta gtccctgaaa actaagaccc
      421 taaccataat cccattgtag acccacctac tcgggtgaat ataaagctcg cacagatcat
      481 ctcacagaca tcagaccttg gcaaaggagc atcctgaaca tctctacaca ctgaggcaac
      541 cctagggccg ccaccatgct gtgttgtatc agaagaacaa agcctgtgga gaagaatgag
      601 gaggctgacc aggaactgca gagcacggta cctagagcta gagaccctcc tgtggctaca
      661 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      721 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      781 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      841 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      901 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      961 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1021 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1081 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1141 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1201 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1261 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1321 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtataagtga
     1381 gaattctgca ggaattgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct
     1441 ggaagttgcc actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt
     1501 gtctgactag gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg
     1561 gcaagttggg aagacaacct gtagggcacg agggggggcc cgaaacccgc tgatcagcct
     1621 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1681 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1741 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1801 attgggaaga caatagcaag gcatgctggg gatgcggtgg gctctatggc aactttatta
     1861 tacaaagttg gcattataaa aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1921 atcagtcaaa ataaaatcat tatttggagc tccatggtag cgttaacgcg gccgcgatat
     1981 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2041 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     2101 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2161 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2221 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2281 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2341 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2401 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2461 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2521 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2581 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2641 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2701 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2761 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     2821 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2881 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2941 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     3001 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     3061 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tttttgccgg atcaagagct
     3181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3241 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3301 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3361 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3421 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3481 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3541 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3601 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3661 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3721 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3781 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3841 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3901 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3961 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     4021 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     4081 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     4141 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     4201 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     4261 ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggccct gca
//
