LOCUS       sequence-193846-        5184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aBb-crystallin promoter fragment in GFP plasmid to assess activity 
            in larval embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5184)
  AUTHORS   Posner M, Murray KL, McDonald MS, Eighinger H, Andrew B, Drossman A,
            Haley Z, Nussbaum J, David LL, Lampi KJ
  TITLE     The zebrafish as a model system for analyzing mammalian and native 
            alpha-crystallin promoter function.
  JOURNAL   PeerJ. 2017 Nov 27;5:e4093. doi: 10.7717/peerj.4093. eCollection 
            2017.
  PUBMED    29201567
REFERENCE   2  (bases 1 to 5184)
  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: "PeerJ."; 
            date: "2017-11-27"; volume: "5:e4093. doi"; pages: " 
            10.7717/peerj.4093. eCollection 2017"
COMMENT     Sequence Label: Danio alpha Bb-crystallin 1kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..5184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             154..948
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             154..948
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(208..227)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     818..837
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1136..1155)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1180..1227
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1556..2144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2045..2064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      3383..3392
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3389..4108
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    4230..4351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4267..4286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4321..4340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4358..4813)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4495..4516)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4707..4726
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4840..4944
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(4946..5184,1..119)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4946..5137
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4969..4989)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(5154..5184,1..105)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
       61 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
      121 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
      181 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
      241 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
      301 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
      361 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
      421 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
      481 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
      541 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
      601 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
      661 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
      721 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
      781 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
      841 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
      901 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
      961 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     1021 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     1081 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     1141 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1201 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     1261 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     1321 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     1381 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     1441 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1501 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1561 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1621 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     1681 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1741 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1801 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1861 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1921 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1981 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2041 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2101 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2161 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2221 cccctgattc tgtggataac cgtattaccg ccatgcatta gttattacta gcgctaccgg
     2281 actcagatct cgagagttca atggtgcgct gtgtgtgtgt gcgcgcgcgc atgtgctttg
     2341 taatgactca tcgtttgtgt gtgactcatc gttgcagaaa ggcttgaatt aactccacaa
     2401 ctaatacatc taatgaccat tgggaaagtt cttacggtag aatttcttac aagcgttacg
     2461 tgagatctgc ttccttcatg tctgtcactg tgctgtttat ctgacacagc caaggcagag
     2521 attgtggcac actctgacag gcgcgtggga acagtgagtg gggagaaaca gcattaaagg
     2581 cacaggccac aaaaacagct ataatgtgtt cagaaaatta caaacttctg aaaggcataa
     2641 caaataatct gattgttgtt ttgagctgaa tttttacaga cagactctgg acacaaagat
     2701 ttatcctaaa ccttgaaaga ttgataaaat aaaagaacat agactgataa cacaccaact
     2761 aagactgctt acctctccaa aatgtcgatg cccattcagt gtgtgccagc tcaagtaagc
     2821 ccagtaatat gttgtccggc tgagggcttg ttcatgggga ttgactttgt atggtggtta
     2881 tcctggcaca gttttctcag ccgcgttcat tgtcagtttc tgtaaagact aaaacctcca
     2941 aaatacacac gggcacatca cctcacgggc atcctaatct ccgttggcat taaatctagc
     3001 cactggaatg ccaagcacct acaaacttaa ctcaatcctg gcaggttcgc cggaccagac
     3061 agcgcagatt ccagtgaaca catgcgggag gcactatcat cacaattaag ggctcacgct
     3121 gtaatccagc cggtatctgc gagaactgaa atccgatcag ctctttggat aaatactgta
     3181 cagagagtga tggagggaag aggggaaggg gagaacacag tgggagggct gcagtgaaag
     3241 aggcaaatga acgtcattca cataattatt ccaaaaaagg tccgtagtgg gctcgtctgc
     3301 caggggccgt cagtatcact ctatttaagt gcgtagacag aagatttgca gaagaggccc
     3361 agactcaaag gatccaccgg tcgccaccat ggtgagcaag ggcgccgagc tgttcaccgg
     3421 catcgtgccc atcctgatcg agctgaatgg cgatgtgaat ggccacaagt tcagcgtgag
     3481 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3541 cggcaagctg cctgtgccct ggcccaccct ggtgaccacc ctgagctacg gcgtgcagtg
     3601 cttctcacgc taccccgatc acatgaagca gcacgacttc ttcaagagcg ccatgcctga
     3661 gggctacatc caggagcgca ccatcttctt cgaggatgac ggcaactaca agtcgcgcgc
     3721 cgaggtgaag ttcgagggcg ataccctggt gaatcgcatc gagctgaccg gcaccgattt
     3781 caaggaggat ggcaacatcc tgggcaataa gatggagtac aactacaacg cccacaatgt
     3841 gtacatcatg accgacaagg ccaagaatgg catcaaggtg aacttcaaga tccgccacaa
     3901 catcgaggat ggcagcgtgc agctggccga ccactaccag cagaataccc ccatcggcga
     3961 tggccctgtg ctgctgcccg ataaccacta cctgtccacc cagagcgccc tgtccaagga
     4021 ccccaacgag aagcgcgatc acatgatcta cttcggcttc gtgaccgccg ccgccatcac
     4081 ccacggcatg gatgagctgt acaagtgagc ggccgcgact ctagatcata atcagccata
     4141 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     4201 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     4261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4321 gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg
     4381 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4441 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4501 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4561 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4621 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4681 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4741 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     4801 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4921 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     4981 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5041 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5101 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5161 ccctaactcc gcccagttcc gccc
//
