LOCUS       sequence-496423-        5089 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  to generate mRNA for membrane mNeonGreen for zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5089)
  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 5089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 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: pAT002
FEATURES             Location/Qualifiers
     source          1..5089
                     /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    948..1069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(959..978)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1013..1032
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1184..1230
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1260..1396
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1441..1512
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1738..2326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1818..1837)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3072..3140,3141..3932)
                     /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(3290..3309)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3945..4081
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4233..4304
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4309..4355)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4373..4421
                     /note="synthetic polyadenylation signal"
     enhancer        4502..4805
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4807..5009
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4960..4980
                     /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 ccatggtcag caagggagag
      181 gaagataaca tggcgagcct gcccgcgacc catgagctgc atatttttgg cagcatcaac
      241 ggtgtggatt tcgatatggt cggccagggc accggtaacc ctaatgatgg ctatgaagag
      301 ctgaacctga aatccaccaa aggtgatctg cagtttagcc cgtggattct ggtcccgcat
      361 atcggctatg gattccatca gtatctgcca tatcctgatg gcatgagccc gttccaggcg
      421 gcgatggtgg atggcagcgg ctaccaggtg catcgcacca tgcagtttga agatggagct
      481 agcctcaccg tgaactaccg ctatacctat gaaggaagcc atattaaagg cgaagctcag
      541 gtcaagggaa ccggctttcc ggctgatggc ccggtcatga caaacagcct gaccgcagct
      601 gattggtgtc gtagcaaaaa aacctacccc aatgataaaa ccattattag cacctttaag
      661 tggagctata ccaccggaaa cggcaaacgt taccgtagca ccgcgcgtac cacctacacc
      721 tttgctaaac cgatggctgc gaactatctg aagaaccagc cgatgtacgt gttccgtaag
      781 accgaactga aacatagcaa gaccgaactg aattttaagg aatggcagaa agcgttcacc
      841 gacgtgatgg gaatggatga actgtacaaa taagaattca tacgtatccg gaaccggtgc
      901 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      961 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1021 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1081 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1141 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1201 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1261 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1321 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1381 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1441 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1501 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1561 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1621 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1681 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1741 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1801 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1861 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1921 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1981 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2041 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2101 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2161 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2221 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2281 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2341 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2401 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2461 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2521 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2581 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2641 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2701 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2761 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2821 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2881 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2941 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     3001 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     3061 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3121 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3181 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3241 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3301 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3361 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3421 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3481 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3541 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3601 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3661 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3721 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3781 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3841 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3901 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3961 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     4021 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     4081 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     4141 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     4201 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     4261 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     4321 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     4381 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     4441 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4501 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4561 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4621 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4681 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4741 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4801 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4861 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4921 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4981 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     5041 cgatcctacc atccactcga cacacccgcc agcggccgct gccaagctt
//
