LOCUS       sequence-10313-e        3311 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 3311)
  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 3311)
  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: pENTRDsRedEx2
FEATURES             Location/Qualifiers
     source          1..3311
                     /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    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      680..689
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             686..1360
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(816..834)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(866..886)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1084..1103
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1278..1301
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    1361..1417
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(1438..1537)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1554..1573)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1555..1573)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1578..1594)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1578..1594)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1696..1715)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1707..2516
                     /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(1774..1793)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2663..3251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3152..3171
                     /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 tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcg ctaccggtcg ccaccatggc ctcctccgag gacgtcatca aggagttcat
      721 gcgcttcaag gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
      781 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggcgg
      841 ccccctgccc ttcgcctggg acatcctgtc cccccagttc cagtacggct ccaaggtgta
      901 cgtgaagcac cccgccgaca tccccgacta caagaagctg tccttccccg agggcttcaa
      961 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1021 cctgcaggac ggctccttca tctacaaggt gaagttcatc ggcgtgaact tcccctccga
     1081 cggccccgta atgcagaaga agactatggg ctgggaggcc tccaccgagc gcctgtaccc
     1141 ccgcgacggc gtgctgaagg gcgagatcca caaggccctg aagctgaagg acggcggcca
     1201 ctacctggtg gagttcaagt ccatctacat ggccaagaag cccgtgcagc tgcccggcta
     1261 ctactacgtg gactccaagc tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1321 gcagtacgag cgcgccgagg gccgccacca cctgttcctg agatctcgag ctcaagcttc
     1381 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatacc
     1441 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     1501 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1561 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1621 tctgatgtta cattgcacaa gataaaataa tatcatcatg aacaataaaa ctgtctgctt
     1681 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     1741 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1801 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1861 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1921 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1981 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2041 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2101 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2161 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2221 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2281 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2341 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2401 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2461 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2521 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     2581 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3301 ttgctcacat g
//
