LOCUS       sequence-497380-        5083 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  to generate mRNA for membrane mStaygold(mBaoJin) for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5083)
  AUTHORS   Tschanz A, Matus DQ, Swinburne I
  TITLE     Systematic comparison of mStayGold and mGold2 variants for live 
            imaging in zebrafish.
  JOURNAL   MicroPubl Biol. 2025. 10.17912/micropub.biology.001847.
REFERENCE   2  (bases 1 to 5083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "MicroPubl 
            Biol. 2025. 10.17912/micropub.biology.001847."
COMMENT     Sequence Label: pAT005
FEATURES             Location/Qualifiers
     source          1..5083
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      25..34
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    942..1063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(953..972)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1007..1026
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1178..1224
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1254..1390
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1435..1506
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1732..2320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1812..1831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3066..3134,3135..3926)
                     /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(3284..3303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3939..4075
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4227..4298
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4303..4349)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4367..4415
                     /note="synthetic polyadenylation signal"
     enhancer        4496..4799
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4801..5003
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4954..4974
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atttaggtga cactatagac tagtgccacc atgggctgca tcaagagcaa gcgtaaggac
       61 aacctgaacg atgatgaagc tgctatggga tgtatcaaat ctaagcgtaa ggataacctt
      121 aatgatgatg agggagcagc tgctcccgct gctgctggat ccatggtaag cacaggagaa
      181 gagctcttca caggtgtggt accattcaag ttccagctga aaggaacgat caatggaaag
      241 tctttcaccg tcgagggcga gggggagggg aattcccacg agggttctca caagggcaag
      301 tatgtgtgta cctccggaaa gctgcctatg tcgtgggcag ctttgggaac ctcctttgga
      361 tatggaatga agtactatac gaagtacccc tctggactga agaactggtt ccgtgaggta
      421 atgcctggag gatttaccta cgatcgtcat atccagtaca aaggtgacgg ttcaatccac
      481 gctaagcacc agcacttcat gaagaacggc acgtaccaca atatcgtcga atttaccggt
      541 caggatttca aggaaaactc cccagtcctg acaggagata tgaacgtcag cctgccaaat
      601 gaagtcccac agatcccccg tgacgatggt gtcgaatgcc cagtgaccct gctctatcct
      661 cttctgtctg ataaatccaa atatgttgaa gctcatcaat acaccatctg caaaccactg
      721 cacaatcaac ctgccccaga cgtcccgtat cattggatcc gcaagcagta tacacaaagt
      781 aaggacgatg cagaagagcg agatcatatc tgccagtccg agactcttga agctcacctt
      841 aagggaatgg atgaactgta caaataagaa ttcatacgta tccggaaccg gtgctagctt
      901 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      961 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     1021 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     1081 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     1141 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     1201 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     1261 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     1321 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1381 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1441 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1501 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1561 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1621 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1681 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1741 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1801 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1861 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1921 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1981 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2041 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2101 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2161 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2221 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2281 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2341 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2401 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2461 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2521 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2581 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2641 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2701 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2761 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2821 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2881 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     2941 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     3001 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     3061 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3121 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3181 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3241 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3301 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3361 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3421 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3481 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3541 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3601 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3661 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3721 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3781 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3841 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3901 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     3961 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     4021 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     4081 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     4141 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     4201 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     4261 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     4321 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     4381 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     4441 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4501 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4561 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4621 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4681 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4741 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4801 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4861 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4921 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4981 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     5041 taccatccac tcgacacacc cgccagcggc cgctgccaag ctt
//
