LOCUS       sequence-270816-        3660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3660)
  AUTHORS   Villefranc JA, Amigo J, Lawson ND
  TITLE     Gateway compatible vectors for analysis of gene function in the 
            zebrafish.
  JOURNAL   Dev Dyn. 2007 Nov . 236(11):3077-87.
  PUBMED    17948311
REFERENCE   2  (bases 1 to 3660)
  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 Dyn. 
            2007 Nov . 236(11):3077-87."
COMMENT     Sequence Label: pENTRbasEGFP
FEATURES             Location/Qualifiers
     source          1..3660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..119
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(278..305)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(397..483)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     532..554
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     546..563
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     547..563
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    579..678
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      755..764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(761..763,764..766,767..1477)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(761..763,764..766,767..1477)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(806..827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1067..1086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1414..1435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1478..1543
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1667..1788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1704..1723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1758..1777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1795..1894)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1911..1930)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1912..1930)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1935..1951)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1935..1951)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2053..2072)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2064..2873
                     /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(2131..2150)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3020..3608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3509..3528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
       61 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      121 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      181 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
      241 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
      301 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
      361 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
      421 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
      481 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
      541 gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac
      601 tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt
      661 tgtacaaaaa agcaggctag actctagagg gtatataatg gatctcgcgt ctcagcctca
      721 ctttgagctc ctccacacgc tagcgctacc ggtcgccacc atggtgagca agggcgagga
      781 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      841 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      901 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      961 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1021 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1081 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1141 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1201 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1261 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1321 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1381 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1441 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ctcgagctca
     1501 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
     1561 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1621 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1681 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1741 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtacccag
     1801 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     1861 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     1921 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1981 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     2041 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     2101 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     2161 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     2221 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     2281 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2341 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     2401 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2461 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2521 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2581 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2641 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2701 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2761 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2821 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2881 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2941 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     3001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
//
