LOCUS       sequence-193848-        8090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish aBb crystallin 4kb promoter segment driving GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8090)
  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 8090)
  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 4kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..8090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      4031..4040
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4037..4756
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    4878..4999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4969..4988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5006..5461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5143..5164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5355..5374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5488..5592
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5594..5951
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5594..5785
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5617..5637)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5802..5937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5986..6780
                     /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             5986..6780
                     /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(6040..6059)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6650..6669
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6968..6987)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7012..7059
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7388..7976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7877..7896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ccgcaccgta caaagatttg gggaaacttg
       61 ctacatgtaa gtctactgaa aatggtggta ttcatatctg tttgacatgc gatcttctga
      121 catgacttta atgtgccagc tgagggcttg aatgtacaga gattatttac aggctggcag
      181 acaaactaaa gagaaccaat tcaaaacagt ttaaagcaga atacatatga cattatacca
      241 tactaggcgg ttttagcaca ttgctaccac tcgtaggaca tcttttatca ttttgctata
      301 atatttttta aatttgttgg tatgtattcc cttaaggtgg catggtggct cagtggttag
      361 cactgttgcc tcacagcaag aaggtcgctg gttcaagtgt cacctggatt agtaggcatt
      421 tctgtgtgga gtttgcatgt tctccccatg tctgcgtggg ttactccagg tacctcgatt
      481 tccttgagta gtaaatgaga gtgtatgggt gttttccagt gccggattgt gggtggaaga
      541 gtatccgctg cataaaacat atgcctgaat agttgccagt tcattcctct ctggcaacct
      601 ctaataaata agggactaag acaattaatt aatgtatttc ccaaggaaaa gtattaagta
      661 atgcaaaata agtatacttt acatataaac atgtatgtcc tatacttgaa tacttcttgg
      721 gactaaactg gcctactttt agtatatttc agtatactct taattttatt atgagtgtaa
      781 cagtagtaca caacatgtct acaatgacct tttacattta ttctacatgt accagcctat
      841 atgcctactg tgcacacagt aataaacata cacactgaaa ctagactcga aactgcaaca
      901 tttggattac aagtcttact ttttaaccgt aaggccacga agtacaataa tatactaata
      961 ctaaactagt agtgttatac tacaagtaac tagtaatttt ctagtgttat actactgcaa
     1021 gtatacttgg caatacttaa gcataaatat tgcataagaa acataaccta cagtatctcc
     1081 taagaaacac ttattgcata aactaacatt atggtggtac ggtaagcttt tatggccgtt
     1141 tctgtcaaaa ggcgtaccaa accataccac tttttcggca ccctttcgaa agggtaccaa
     1201 acacgagtaa gggtaccaga aggcggaacc agacgtgcag ctgcacacta cagagatacg
     1261 tcattcgctc aggcaacaag ccagaatgaa aacaaaggaa ccgacatgtt ttaaatattc
     1321 agcagagaga ttacaccata attatatata catataataa caagccatag tcgacccggg
     1381 ctcaaacaaa ccttgtcgtc gtcatgatga acagccacaa agccaaaaag aagagcagat
     1441 tttacgctgt gctccttagt tttttacgag ccagtctgaa gcacaagcgg tttcgctttc
     1501 tagctttcgc ttgccgtgca tttatatttg aaataactaa tttcctgagc tcatgataat
     1561 aacgtgcgct tgattattga tgtgcttttg aaacccgatc ctgtcagaaa ctgacaaacg
     1621 caagagtgac acgaaaaaca aagaagaagc cggaaaaaaa ggagcacatt attctttcag
     1681 caaacgtgaa caaactgcca tgtttaacca ttattatcac ctttttgact attatgaact
     1741 cggaatgata tcattatttt ctaacgagag gctatacgtg ctgctaaaga tacatttaat
     1801 gatgagaggt ttgcactcac tgtggggtat atttcgtgtt gttttgaacc taaattagga
     1861 ctaaatgtat gctgtttgta gtttttctgt aggtggtaac atatcagaga ctgtaagagt
     1921 ctgtatgtgt tcatatatgt tatttatttt aagtaattgc agacgttaca gtgggctatt
     1981 tagcactgtc attaatctgc agttataatc aaattatgtt tatagaaaag ttagtaataa
     2041 acatttatac acaagtattt atgtgtataa agcatctgtt ttgtgagaag tgcttctcat
     2101 atgatatgtg agcgacccgt atagctttac tgttgacatt ttaacctttt aaactttgtc
     2161 gtacatcacg cccaccaaaa gagtaccatt ggtacccttt tagcattgga aacgcaagcc
     2221 taataagatg actcataccg acccgtgccg taccgtactg aaccaggcag tagaaacaag
     2281 ccattagaaa cctttttcaa gaacataatg ataattaatg ctgatgataa ttaaatgaat
     2341 tgcaaaaaac tgaaaaataa atacataaat aaataaaaaa acactctcct tcactattgt
     2401 taaagctgtt tggacagaac tgtgtgtagg tatagtgtgt ccactgtcat attgcagtga
     2461 tataaacaag tctctttttt tatttcctga cgttaaaata ggactcaaat ccctcccatt
     2521 ttgaggccca ctgcaatgtg acataggagt gcggtttcca caaccactga actgattgat
     2581 agctgcatat taaaatatct gtaactcgta taatcatatc aacaagacag gacgtgcgta
     2641 aagcaactgg aattaaaaga tctgttcagc ttgtgtgatc atcaatcatc attaaaagtg
     2701 atcaaggagg gggtttacaa gtttaaaaag tttttaaaac agtgcatgtt tgtaataaat
     2761 cacagtgatt tacttcagct ttactttatc accacagctg catgtcagta ctattataaa
     2821 agaggatgct tcaatcccag tttgtggata ttaaatcagg tgtattttgt acattaacat
     2881 aatggatatc catacagcag tggatattac catgtatcct gtcacatttg tgtgcaaaaa
     2941 tttgcaaagt tcaatggtgc gctgtgtgtg tgtgcgcgcg cgcatgtgct ttgtaatgac
     3001 tcatcgtttg tgtgtgactc atcgttgcag aaaggcttga attaactcca caactaatac
     3061 atctaatgac cattgggaaa gttcttacgg tagaatttct tacaagcgtt acgtgagatc
     3121 tgcttccttc atgtctgtca ctgtgctgtt tatctgacac agccaaggca gagattgtgg
     3181 cacactctga caggcgcgtg ggaacagtga gtggggagaa acagcattaa aggcacaggc
     3241 cacaaaaaca gctataatgt gttcagaaaa ttacaaactt ctgaaaggca taacaaataa
     3301 tctgattgtt gttttgagct gaatttttac agacagactc tggacacaaa gatttatcct
     3361 aaaccttgaa agattgataa aataaaagaa catagactga taacacacca actaagactg
     3421 cttacctctc caaaatgtcg atgcccattc agtgtgtgcc agctcaagta agcccagtaa
     3481 tatgttgtcc ggctgagggc ttgttcatgg ggattgactt tgtatggtgg ttatcctggc
     3541 acagttttct cagccgcgtt cattgtcagt ttctgtaaag actaaaacct ccaaaataca
     3601 cacgggcaca tcacctcacg ggcatcctaa tctccgttgg cattaaatct agccactgga
     3661 atgccaagca cctacaaact taactcaatc ctggcaggtt cgccggacca gacagcgcag
     3721 attccagtga acacatgcgg gaggcactat catcacaatt aagggctcac gctgtaatcc
     3781 agccggtatc tgcgagaact gaaatccgat cagctctttg gataaatact gtacagagag
     3841 tgatggaggg aagaggggaa ggggagaaca cagtgggagg gctgcagtga aagaggcaaa
     3901 tgaacgtcat tcacataatt attccaaaaa aggtccgtag tgggctcgtc tgccaggggc
     3961 cgtcagtatc actctattta agtgcgtaga cagaagattt gcagaagagg cccagactca
     4021 aacaccggtc gccaccatgg tgagcaaggg cgccgagctg ttcaccggca tcgtgcccat
     4081 cctgatcgag ctgaatggcg atgtgaatgg ccacaagttc agcgtgagcg gcgagggcga
     4141 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4201 tgtgccctgg cccaccctgg tgaccaccct gagctacggc gtgcagtgct tctcacgcta
     4261 ccccgatcac atgaagcagc acgacttctt caagagcgcc atgcctgagg gctacatcca
     4321 ggagcgcacc atcttcttcg aggatgacgg caactacaag tcgcgcgccg aggtgaagtt
     4381 cgagggcgat accctggtga atcgcatcga gctgaccggc accgatttca aggaggatgg
     4441 caacatcctg ggcaataaga tggagtacaa ctacaacgcc cacaatgtgt acatcatgac
     4501 cgacaaggcc aagaatggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggatgg
     4561 cagcgtgcag ctggccgacc actaccagca gaataccccc atcggcgatg gccctgtgct
     4621 gctgcccgat aaccactacc tgtccaccca gagcgccctg tccaaggacc ccaacgagaa
     4681 gcgcgatcac atgatctact tcggcttcgt gaccgccgcc gccatcaccc acggcatgga
     4741 tgagctgtac aagtgagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     4801 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     4861 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4921 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4981 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     5041 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     5101 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     5161 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     5221 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     5281 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     5341 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5401 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     5461 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5521 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5581 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     5641 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5701 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5761 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     5821 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     5881 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     5941 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     6001 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     6061 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     6121 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     6181 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     6241 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     6301 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     6361 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     6421 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     6481 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     6541 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     6601 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     6661 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     6721 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     6781 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     6841 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     6901 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     6961 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     7021 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     7081 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     7141 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     7201 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     7261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
//
