LOCUS       sequence-193849-        9079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish aBb crystallin 5kb promoter segment driving GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9079)
  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 9079)
  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: Zebrafish aBb crystallin 5kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..9079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      5020..5029
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5026..5745
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    5867..5988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5904..5923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5958..5977
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5995..6450)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6132..6153)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6344..6363
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6477..6581
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6583..6940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6583..6774
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6606..6626)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      6791..6926
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6975..7769
                     /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             6975..7769
                     /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(7029..7048)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7639..7658
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7957..7976)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8001..8048
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8377..8965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8866..8885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctatttagac ctgcttttag ttggtcaatg
       61 gtgcagtcta tttcagttcc tcaaaacagc aatgccaaca atgccttaac acacctccat
      121 tctagaccgg cacacccatg agtccacaaa gtggcgcaaa aaacgtgaaa attagggttg
      181 cgttggtctg aaaatagcaa caaatcacgt caaacacgtc ttgcgcctta ttgtgctgag
      241 tgtatagggc cctttaagtt tgaagtagtt ttcagtttcc aaagctcaaa ttttaatagc
      301 tttaagactc agattacagc attattagag cattctgtat cacatcatta gccattttca
      361 atgattttat gggtggtcct taaagtgttg ctaggtgatt aatatggcag tatctgaaat
      421 cagcccctat tcccctaaac tgtgcactat ttgacagagc agccattttt agtggtgtct
      481 gaaaccatag tggacttttt gaagtgcact caatcaatcc taaaatgcac cataacagta
      541 cgtttacaac tcacatgcaa gaaaattccc ataatgcact gagtgagtct gcattgcgaa
      601 taaatttcga tgtctgacca caaaacggcg tccacagcag aatcaaagtt tctaaataat
      661 tgattattta attaaagtaa cctttgtata tatgatagga atcaaaaaac tttgttttgc
      721 catcattgtc agctcagtaa gtgttaaata gctatcacag tgtctgacta agaagttaaa
      781 gaagtaaatt ctaatctgtc tttattaact agcaaaacaa aaacaaaaga gcccactgct
      841 ctaattcact ccatttaatt aatttattga agtggactat attactggac tcagggaaat
      901 atgaaagggg tacaaggggt gattttggat acagtatata tgttagcata gctagcatgc
      961 tttagctgta atctacacag tagattgcag gagtaaacgg cacaaacact attcataaac
     1021 cgcaccgaac aaagatttgg ggaaacttgc tacatgtaag tctactgaaa atggtggtat
     1081 tcatatctgt ttgacatgcg atcttctgac atgactttaa tgtgccagct gagggcttga
     1141 atgtacagag attatttaca ggctggcaga caaactaaag agaaccaatt caaaacagtt
     1201 taaagcagaa tacatatgac attataccat actaggcggt tttagcacat tgctaccact
     1261 cgtaggacat cttttatcat tttgctataa tattttttaa atttgttggt atgtattccc
     1321 ttaaggtggc atggtggctc agtggttagc actgttgcct cacagcaaga aggtcgctgg
     1381 ttcaagtgtc acctggatta gtaggcattt ctgtgtggag tttgcatgtt ctccccatgt
     1441 ctgcgtgggt tactccaggt acctcgattt ccttgagtag taaatgagag tgtatgggtg
     1501 ttttccagtg ccggattgtg ggtggaagag tatccgctgc ataaaacata tgcctgaata
     1561 gttgccagtt cattcctctc tggcaacctc taataaataa gggactaaga caattaatta
     1621 atgtatttcc caaggaaaag tattaagtaa tgcaaaataa gtatacttta catataaaca
     1681 tgtatgtcct atacttgaat acttcttggg actaaactgg cctactttta gtatatttca
     1741 gtatactctt aattttatta tgagtgtaac agtagtacac aacatgtcta caatgacctt
     1801 ttacatttat tctacatgta ccagcctata tgcctactgt gcacacagta ataaacatac
     1861 acactgaaac tagactcgaa actgcaacat ttggattaca agtcttactt tttaaccgta
     1921 aggccacgaa gtacaataat atactaatac taaactagta gtgttatact acaagtaact
     1981 agtaattttc tagtgttata ctactgcaag tatacttggc aatacttaag cataaatatt
     2041 gcataagaaa cataacctac agtatctcct aagaaacact tattgcataa actaacatta
     2101 tggtggtacg gtaagctttt atggccgttt ctgtcaaaag gcgtaccaaa ccataccact
     2161 ttttcggcac cctttcgaaa gggtaccaaa cacgagtaag ggtaccagaa ggcggaacca
     2221 gacgtgcagc tgcacactac agagatacgt cattcgctca ggcaacaagc cagaatgaaa
     2281 acaaaggaac cgacatgttt taaatattca gcagagagat tacaccataa ttatatatac
     2341 atataataac aagccatagt cgacccgggc tcaaacaaac cttgtcgtcg tcatgatgaa
     2401 cagccacaaa gccaaaaaga agagcagatt ttacgctgtg ctccttagtt ttttacgagc
     2461 cagtctgaag cacaagcggt ttcgctttct agctttcgct tgccgtgcat ttatatttga
     2521 aataactaat ttcctgagct catgataata acgtgcgctt gattattgat gtgcttttga
     2581 aacccgatcc tgtcagaaac tgacaaacgc aagagtgaca cgaaaaacaa agaagaagcc
     2641 ggaaaaaaag gagcacatta ttctttcagc aaacgtgaac aaactgccat gtttaaccat
     2701 tattatcacc tttttgacta ttatgaactc ggaatgatat cattattttc taacgagagg
     2761 ctatacgtgc tgctaaagat acatttaatg atgagaggtt tgcactcact gtggggtata
     2821 tttcgtgttg ttttgaacct aaattaggac taaatgtatg ctgtttgtag tttttctgta
     2881 ggtggtaaca tatcagagac tgtaagagtc tgtatgtgtt catatatgtt atttatttta
     2941 agtaattgca gacgttacag tgggctattt agcactgtca ttaatctgca gttataatca
     3001 aattatgttt atagaaaagt tagtaataaa catttataca caagtattta tgtgtataaa
     3061 gcatctgttt tgtgagaagt gcttctcata tgatatgtga gcgacccgta tagctttact
     3121 gttgacattt taacctttta aactttgtcg tacatcacgc ccaccaaaag agtaccattg
     3181 gtaccctttt agcattggaa acgcaagcct aataagatga ctcataccga cccgtgccgt
     3241 accgtactga accaggcagt agaaacaagc cattagaaac ctttttcaag aacataatga
     3301 taattaatgc tgatgataat taaatgaatt gcaaaaaact gaaaaataaa tacataaata
     3361 aataaaaaaa cactctcctt cactattgtt aaagctgttt ggacagaact gtgtgtaggt
     3421 atagtgtgtc cactgtcata ttgcagtgat ataaacaagt ctcttttttt atttcctgac
     3481 gttaaaatag gactcaaatc cctcccattt tgaggcccac tgcaatgtga cataggagtg
     3541 cggtttccac aaccactgaa ctgattgata gctgcatatt aaaatatctg taactcgtat
     3601 aatcatatca acaagacagg acgtgcgtaa agcaactgga attaaaagat ctgttcagct
     3661 tgtgtgatca tcaatcatca ttaaaagtga tcaaggaggg ggtttacaag tttaaaaagt
     3721 ttttaaaaca gtgcatgttt gtaataaatc acagtgattt acttcagctt tactttatca
     3781 ccacagctgc atgtcagtac tattataaaa gaggatgctt caatcccagt ttgtggatat
     3841 taaatcaggt gtattttgta cattaacata atggatatcc atacagcagt ggatattacc
     3901 atgtatcctg tcacatttgt gtgcaaaaat ttgcaaagtt caatggtgcg ctgtgtgtgt
     3961 gtgcgcgcgc gcatgtgctt tgtaatgact catcgtttgt gtgtgactca tcgttgcaga
     4021 aaggcttgaa ttaactccac aactaataca tctaatgacc attgggaaag ttcttacggt
     4081 agaatttctt acaagcgtta cgtgagatct gcttccttca tgtctgtcac tgtgctgttt
     4141 atctgacaca gccaaggcag agattgtggc acactctgac aggcgcgtgg gaacagtgag
     4201 tggggagaaa cagcattaaa ggcacaggcc acaaaaacag ctataatgtg ttcagaaaat
     4261 tacaaacttc tgaaaggcat aacaaataat ctgattgttg ttttgagctg aatttttaca
     4321 gacagactct ggacacaaag atttatccta aaccttgaaa gattgataaa ataaaagaac
     4381 atagactgat aacacaccaa ctaagactgc ttacctctcc aaaatgtcga tgcccattca
     4441 gtgtgtgcca gctcaagtaa gcccagtaat atgttgtccg gctgagggct tgttcatggg
     4501 gattgacttt gtatggtggt tatcctggca cagttttctc agccgcgttc attgtcagtt
     4561 tctgtaaaga ctaaaacctc caaaatacac acgggcacat cacctcacgg gcatcctaat
     4621 ctccgttggc attaaatcta gccactggaa tgccaagcac ctacaaactt aactcaatcc
     4681 tggcaggttc gccggaccag acagcgcaga ttccagtgaa cacatgcggg aggcactatc
     4741 atcacaatta agggctcacg ctgtaatcca gccggtatct gcgagaactg aaatccgatc
     4801 agctctttgg ataaatactg tacagagagt gatggaggga agaggggaag gggagaacac
     4861 agtgggaggg ctgcagtgaa agaggcaaat gaacgtcatt cacataatta ttccaaaaaa
     4921 ggtccgtagt gggctcgtct gccaggggcc gtcagtatca ctctatttaa gtgcgtagac
     4981 agaagatttg cagaagaggc ccagactcaa acaccggtcg ccaccatggt gagcaagggc
     5041 gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc tgaatggcga tgtgaatggc
     5101 cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     5161 aagttcatct gcaccaccgg caagctgcct gtgccctggc ccaccctggt gaccaccctg
     5221 agctacggcg tgcagtgctt ctcacgctac cccgatcaca tgaagcagca cgacttcttc
     5281 aagagcgcca tgcctgaggg ctacatccag gagcgcacca tcttcttcga ggatgacggc
     5341 aactacaagt cgcgcgccga ggtgaagttc gagggcgata ccctggtgaa tcgcatcgag
     5401 ctgaccggca ccgatttcaa ggaggatggc aacatcctgg gcaataagat ggagtacaac
     5461 tacaacgccc acaatgtgta catcatgacc gacaaggcca agaatggcat caaggtgaac
     5521 ttcaagatcc gccacaacat cgaggatggc agcgtgcagc tggccgacca ctaccagcag
     5581 aataccccca tcggcgatgg ccctgtgctg ctgcccgata accactacct gtccacccag
     5641 agcgccctgt ccaaggaccc caacgagaag cgcgatcaca tgatctactt cggcttcgtg
     5701 accgccgccg ccatcaccca cggcatggat gagctgtaca agtgagcggc cgcgactcta
     5761 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5821 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5881 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5941 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     6001 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     6061 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6121 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6181 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6241 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     6301 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6361 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6421 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     6481 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     6541 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     6601 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     6661 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     6721 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     6781 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     6841 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     6901 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     6961 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     7021 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     7081 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     7141 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     7201 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     7261 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     7321 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     7381 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     7441 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     7501 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     7561 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     7621 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     7681 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     7741 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7801 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     7861 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7921 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     7981 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     8041 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     8101 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     8161 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     8221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     8281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     8341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     8401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     8461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     8521 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     8581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     8641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     8701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     9001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     9061 ccgtattacc gccatgcat
//
