LOCUS       sequence-385226-        3785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway p3E 2A fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3785)
  AUTHORS   Kirchgeorg L, Felker A, van Oostrom M, Chiavacci E, Mosimann C
  TITLE     Cre/lox-controlled spatiotemporal perturbation of FGF signaling in 
            zebrafish.
  JOURNAL   Dev Dyn. 2018 Oct;247(10):1146-1159. doi: 10.1002/dvdy.24668.
  PUBMED    30194800
REFERENCE   2  (bases 1 to 3785)
  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"; doi: "10.1002/dvdy"; 
            journalName: "Dev Dyn"; date: "2018-10"; volume: "247"; issue: "10";
            pages: "1146-1159"
COMMENT     Sequence Label: pAF018 p3E-2A-mCerulean-CAAX
FEATURES             Location/Qualifiers
     source          1..3785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(61..184)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             196..252
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(262..264,265..267,268..978)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(307..328)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(568..587)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     915..936
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1087..1105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1110..1244
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1134..1153)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1188..1207
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(1363..1458)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1491..1509)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1514..1530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1514..1530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1632..1651)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1643..2452
                     /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(1710..1729)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2599..3187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3088..3107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3341..3358
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3517..3544)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3636..3722)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3771..3785,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3785,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aaaattctac aggtcactaa taccatctaa gtagttggtt
       61 catagtgact gcatatgttg tgttttacag tattatgtag tctgtttttt atgcaaaatc
      121 taatttaata tattgatatt tatatcattt tacgtttctc gttcaacttt cttgtacaaa
      181 gtggggggat ctggagccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa
      241 aaccccggtc ctagggatcc catggtgagc aagggcgagg agctgttcac cggggtggtg
      301 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      361 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      421 ctgcccgtgc cctggcccac cctcgtgacc accctgacct ggggcgtgca gtgcttcgcc
      481 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      541 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      601 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      661 gacggcaaca tcctggggca caagctggag tacaacgcca tcagcgacaa cgtctatatc
      721 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
      781 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      841 gtgctgctgc ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac
      901 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      961 atggacgagc tgtacaaggg gcgcgccaag ctgaaccctc ctgatgagag tggccccggc
     1021 tgcatgagct gcaagtgtgt gctctcctga gaattcaagg cctctcgagc ctctagattc
     1081 tgcagcccta tagtgagtcg tattacgtag atccagacat gataagatac attgatgagt
     1141 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1201 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1261 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc gcggccgcgg
     1321 cgccaatgca ttgggcccgg taccgatcca ccggatcttg accaacttta ttatacaaag
     1381 ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     1441 aaaataaaat cattatttgg agctccatgg tagcgttaac gcggccatcc cctatagtga
     1501 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     1561 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     1621 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
     1681 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
     1741 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
     1801 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
     1861 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
     1921 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
     1981 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
     2041 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
     2101 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
     2161 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
     2221 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     2281 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     2341 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     2401 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     2461 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     2521 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     2581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2701 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2761 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2821 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2881 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2941 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3001 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3061 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3121 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3181 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3241 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     3301 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     3361 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     3421 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     3481 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     3541 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     3601 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     3661 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     3721 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
     3781 acgtt
//
