LOCUS       sequence-193845-        7099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  aBA-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 7099)
  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 7099)
  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 Ba-crystallin 3kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3040..3049
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3046..3765
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    3887..4008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3924..3943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3978..3997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4015..4470)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4152..4173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4364..4383
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4497..4601
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4603..4960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4603..4794
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4626..4646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4811..4946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4995..5789
                     /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             4995..5789
                     /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(5049..5068)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5659..5678
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5977..5996)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6021..6068
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6397..6985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6886..6905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat cgaaaaaaaa aaaagaaaga aagaaaagaa
       61 agaaaagaaa atatctagtg agtggctttt atgttgaaga aaatgccttg ttgatgccag
      121 tattctggct tgggatgtct ggttcaaact gatacaaagg caatagtaac tcaaaaaaca
      181 aaataacttt tatagaggtc tgaagcatct atgatcacaa aacacattaa aactgagcta
      241 aagcaataat agtaggaaga agagcacact aatgacactc ctgtctgaaa acaacaacaa
      301 actgaggcta cagttcagac atatttacag ctaacaaatc tgcagcaacc gagtgacatc
      361 aagctgatat ggactaacaa ccctgtggga tgtttatagc aacttcttga atatgctgcg
      421 caaataatta aaactattta aggcaaaaga gggtccaacc tgttctagca ttgtttacat
      481 aaaggggtat ttcaaccaaa aatgaaaatt accttataat ttattttccc tcatgtgatt
      541 ttaagccttt atgagtttct ttcttctgtt gaacacaaaa ggaggtattt tggaaaatgc
      601 tggctcgacc catcgacttc caaaaagact gaaattcaac tgctgcatcc aaattttatc
      661 tgtcctagta gtagtagtaa aaatagtaaa gtagtaaagt aaagtaaata gtaatagtaa
      721 aaagagttaa caaactacaa acatggcaga cgcaagacct accattaatc ctttggaaat
      781 gaattgtttg aatgaccaat aatttatatc taacctactg gaaaatattt gaatatatat
      841 atatatatat taaaaaaagc ttggctctta ccttacaaat tttgtgttcc caacaaagtt
      901 aaagaattac gtttgaaagt gttacacaat atctacccca caaatgagat tgcatctaaa
      961 ttcggttatc tcaaattgtg tcttctgtaa ctcaagcatc gaatcgattg atcatctttt
     1021 tgtcaaatgc ccacatgtta atgtgttgtg gtctgatttt ctgtctattt ttttatctaa
     1081 atgtaagttt aacattatct ttttccatgt gtgatataat ttgttacttt tcccatgaca
     1141 aacataacat ataatttata ggcaatttat ttattttatt gaataaatat tttattcaca
     1201 aatgtaaatt ttcttcatca ttacctaggt tgcagaattg ttttgtgact ttgctttaaa
     1261 ttgtttaatt gctacttcac aaaatgttta cagcataaaa aagtgttaaa tttcattcat
     1321 tcattcattc attttcttgt cggcttagtc cctttattaa tctggggtcg ccgcagtgga
     1381 atgaactgcc aacttatcca gctcattttt acgcagcgga tgcccttcca gccgcaaccc
     1441 atctctggaa aaaaatgtta aaattgtgaa attttaaaag gactttgtaa atattattta
     1501 attttataat ttgcctctgt actctgtttt ttattatctt attttattat ttttttatta
     1561 ttattaattt tacttgttta ttattgtact tgatttatta ttgtacattg ttatgattaa
     1621 tatgtttgta ttgttaaaac acaataaaaa aaaaaagagt ttgtgaggtt aatgtgcagc
     1681 aacaatgtga acctttataa aagtgtgttt ggccgttaac ttttgaatgt gtaattccga
     1741 aaagcatctg aagagatttc tctgcatgaa acacacataa atggcagatc aacctgctga
     1801 tgcttcttca attcggtagg agattaaata tgagagaaat gtggttgatg tgtggagatt
     1861 attgacagtg catgtaacta agcagcataa tggtttgtca cagaacgtgc atagtcttgt
     1921 tacacactca aaagtgtgca ccttttttca taaagcaatg aatgcgataa catttttcaa
     1981 aatatctttt ttttgtgcta aggttttgaa caagtgattg gttagtaaat gatgacttta
     2041 ataaaaaaaa gatggctact ggattgatgg atggacaaga cagaccaaac acctgaaatt
     2101 gtgtttccat ccccatttca acactttaca ttaggtggcc tgaactagta tgtacttgca
     2161 ttaaaaaaat aaatataatg tacttactgt gttcataatg tattgccgaa cacttctgat
     2221 gctcttgagg tgggatacgg gtacggttag gaacaggttt ggtggtttga gtaagtttaa
     2281 gggtgggtaa aggtgtaagg gatggtcaac catgtagtta catcaatgaa ttacagaggt
     2341 agttgcatga atgtatttaa tcaatcataa gtacatgtgt ttactcaata attacaatgt
     2401 aacaaatgtt taatttcaat gtaaatacat attagtaaag gccaccttat ataaagtggg
     2461 acccaactgc ctatcttcct aatgttccta agacgggaca gcaggagtcc tcttgggctc
     2521 attagtcatg tcagatttac agcagatcat cagtcttgct gattcctgca gtcccggcca
     2581 ttttagggca taatattatc tttctgcctg agttcttctg tgatagcagt caggatgggg
     2641 cagataatat ctgtccgcat cccgcccagg gcatggggtg cagaatccct cctggcacaa
     2701 gagcttgctg ggatgagtcc agtttacatg gcggcttcac tcagaaccat tctaacttag
     2761 ccaggcaaat atttttccag gcttgtgagc agaggggcat taaaactatt cagagcccag
     2821 catttgagac tctatgatct aaaagagagt gaagggtgat tcaataggtt tttgtttgaa
     2881 tgagccagag gggaagaggg cagcaaaaaa cactttacct cataactgga gatagatcag
     2941 tggctgacac gaggtttttg ggatggggca taaatacagg gcccctgcag aacctgcagc
     3001 agatccgctg tcagctccaa actaaggtac acaccggtcg ccaccatggt gagcaagggc
     3061 gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc tgaatggcga tgtgaatggc
     3121 cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3181 aagttcatct gcaccaccgg caagctgcct gtgccctggc ccaccctggt gaccaccctg
     3241 agctacggcg tgcagtgctt ctcacgctac cccgatcaca tgaagcagca cgacttcttc
     3301 aagagcgcca tgcctgaggg ctacatccag gagcgcacca tcttcttcga ggatgacggc
     3361 aactacaagt cgcgcgccga ggtgaagttc gagggcgata ccctggtgaa tcgcatcgag
     3421 ctgaccggca ccgatttcaa ggaggatggc aacatcctgg gcaataagat ggagtacaac
     3481 tacaacgccc acaatgtgta catcatgacc gacaaggcca agaatggcat caaggtgaac
     3541 ttcaagatcc gccacaacat cgaggatggc agcgtgcagc tggccgacca ctaccagcag
     3601 aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct gtccacccag
     3661 agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt cggcttcgtg
     3721 accgccgccg ccatcaccca cggcatggat gagctgtaca agtgagcggc cgcgactcta
     3781 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     3841 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     3901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     4021 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4081 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4141 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4201 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4261 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4321 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4381 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4441 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     4501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     4621 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     4681 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     4741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     4801 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4861 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4921 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4981 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5041 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     5101 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     5161 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     5221 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     5281 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     5341 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     5401 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     5461 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     5521 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     5581 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     5641 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     5701 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     5761 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     5821 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     5881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     5941 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     6001 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     6061 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     6121 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     6181 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     6241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     7081 ccgtattacc gccatgcat
//
