LOCUS       sequence-398283-        5811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector for the expression of a fluorescent fusion of 
            N-ter emiRFP703 and a protein of interest into embryos, by mRNA 
            microinjection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  TITLE     Lemaire Lab Plasmids 2022
REFERENCE   2  (bases 1 to 5811)
  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"
COMMENT     Sequence Label: pDEST-Spe4-Kozak-emiRFP703-RfA
FEATURES             Location/Qualifiers
     source          1..5811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(126..194,195..986)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(344..363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1157..1745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1646..1665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1899..1916
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2033..2054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2069..2074,2075..2092,2093..2099)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2107..2123
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2112..2134
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2131..2147
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2131..2147
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2168..2186
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           2211..2253
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     protein_bind    3205..3329
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3354..3359,3360..3377,3378..3384)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3438..4097
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3504..3523)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4439..4744
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4623..4642
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4785..4909)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     3'UTR           4954..5079
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(5167..5186)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5168..5186)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5193..5210)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5193..5209)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5202..5224)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5350..5805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5487..5508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5699..5718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
      181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
      241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
      301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
      361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
      421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
      481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
      541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
      601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
      661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
      721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
      781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
      841 gatctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
      901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
      961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     1081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     1141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     1201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     1261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     1321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     1381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     1441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     1501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     1561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     1621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     1681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     1741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     1801 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     1861 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     1921 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1981 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     2041 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     2101 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     2161 agctcgaaat taaccctcac taaagggaac aaaagctgga gctaagcttg cttgttcttt
     2221 ttgcagaagc tcagaataaa cgctcaactt tggcagatct cagaaaaaat ggcggaaggc
     2281 tccgtcgcca ggcagcctga cctcttgacc tgcgaacatg aagagatcca cctcgccggc
     2341 tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac atgatcatcg cgtcatccag
     2401 gccagcgcca acgccgcgga atttctgaat ctcggaagcg tactcggcgt tccgctcgcc
     2461 gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc tcgatcccac cgccgaaggc
     2521 atgccggtcg cggtgcgctg ccggatcggc aatccctcta cggagtactg cggtctgatg
     2581 catcggcctc cggaaggcgg gctgatcatc gaactcgaac gtgccggccc gtcgatcgat
     2641 ctgtcaggca cgctggcgcc ggcgctggag cggatccgca cggcgggttc actgcgcgcg
     2701 ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg gctacgaccg ggtgatggtg
     2761 tatcgtttcg atgagcaagg ccacggcctg gtattctccg agtgccatgt gcctgggctc
     2821 gaatcctatt tcggcaaccg ctatccgtcg tcgctggtcc cgcagatggc gcggcagctg
     2881 tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct atcagccggt gccgctggag
     2941 ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt cgggctgctt cctgcgctcg
     3001 atgtcgccga tccatctgca gttcctgaag gacatgggcg tgcgcgccac cctggcggtg
     3061 tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct gtcaccatta tctgccgcgc
     3121 ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg ccgaaaggat cgcgacgcgg
     3181 atcaccgcgc ttgagagcgt aatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa
     3241 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac
     3301 tgtaaaacac aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac
     3361 tttatgcttc cggctcgtat aatgtgtgga ttttgagtta ggatccggcg agattttcag
     3421 gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc
     3481 aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc
     3541 agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt
     3601 tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta
     3661 tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt
     3721 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
     3781 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
     3841 ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca
     3901 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
     3961 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
     4021 tctgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
     4081 ggcagggcgg ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt
     4141 atttgcgcgc tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt
     4201 caaaaagagg tgtgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca
     4261 gttgctcaag gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat
     4321 gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag
     4381 gtcgcccggt ttattgaaat gaacggctct tttgctgacg agaacaggga ctggtgaaat
     4441 gcagtttaag gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca
     4501 gagtgatatt attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct
     4561 gctgtcagat aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg
     4621 gcgcatgatg accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc
     4681 tgatctcagc caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat
     4741 ataaatgtca ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat
     4801 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga
     4861 tatttatatc attttacgtt tctcgttcag ctttcttgta caaagtggtg ataaacttag
     4921 atatcgcggc cgcggaagcg ctggttacct aggaaccagc ctcaagaaca cccgaatgga
     4981 gtctctaagc tacataatac caacttacac tttacaaaat gttgtccccc aaaatgtagc
     5041 cattcgtatc tgctcctaat aaaaagaaag tttcttcaca ttctaaaaaa aaaaaaaaaa
     5101 aaaaaaaaaa aaaaaaamcc cccccccccc ccccctgcag ggccaagtcg gccggcgcgc
     5161 cattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
     5221 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     5281 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     5341 ggcgaatgga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     5401 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     5461 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     5521 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     5581 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     5641 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     5701 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     5761 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt g
//
