LOCUS       sequence-193847-        6169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zebrafish aBb crystallin 2kb promoter segment driving GFP 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  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 6169)
  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 2kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..6169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2110..2119
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2116..2835
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2957..3078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2994..3013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3048..3067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3085..3540)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3222..3243)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3434..3453
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3567..3671
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3673..4030
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3673..3864
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3696..3716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3881..4016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4065..4859
                     /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             4065..4859
                     /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(4119..4138)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4729..4748
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5047..5066)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5091..5138
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5467..6055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5956..5975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagagac gttacagtgg gctatttagc
       61 actgtcatta atctgcagtt ataatcaaat tatgtttata gaaaagttag taataaacat
      121 ttatacacaa gtatttatgt gtataaagca tctgttttgt gagaagtgct tctcatatga
      181 tatgtgagcg acccgtatag ctttactgtt gacattttaa ccttttaaac tttgtcgtac
      241 atcacgccca ccaaaagagt accattggta cccttttagc attggaaacg caagcctaat
      301 aagatgactc ataccgaccc gtgccgtacc gtactgaacc aggcagtaga aacaagccat
      361 tagaaacctt tttcaagaac ataatgataa ttaatgctga tgataattaa atgaattgca
      421 aaaaactgaa aaataaatac ataaataaat aaaaaaacac tctccttcac tattgttaaa
      481 gctgtttgga cagaactgtg tgtaggtata gtgtgtccac tgtcatattg cagtgatata
      541 aacaagtctc tttttttatt tcctgacgtt aaaataggac tcaaatccct cccattttga
      601 ggcccactgc aatgtgacat aggagtgcgg tttccacaac cactgaactg attgatagct
      661 gcatattaaa atatctgtaa ctcgtataat catatcaaca agacaggacg tgcgtaaagc
      721 aactggaatt aaaagatctg ttcagcttgt gtgatcatca atcatcatta aaagtgatca
      781 aggagggggt ttacaagttt aaaaagtttt taaaacagtg catgtttgta ataaatcaca
      841 gtgatttact tcagctttac tttatcacca cagctgcatg tcagtactat tataaaagag
      901 gatgcttcaa tcccagtttg tggatattaa atcaggtgta ttttgtacat taacataatg
      961 gatatccata cagcagtgga tattaccatg tatcctgtca catttgtgtg caaaaatttg
     1021 caaagttcaa tggtgcgctg tgtgtgtgcg cgcgcgcatg tgctttgtaa tgactcatcg
     1081 tttgtgtgtg actcatcgtt gcagaaaggc ttgaattaac tccacaacta atacatctaa
     1141 tgaccattgg gaaagttctt acggtagaat ttcttacaag cgttacgtga gatctgcttc
     1201 cttcatgtct gtcactgtgc tgtttatctg acacagccaa ggcagagatt gtggcacact
     1261 ctgacaggcg cgtgggaaca gtgagtgggg agaaacagca ttaaaggcac aggccacaaa
     1321 aacagctata atgtgttcag aaaattacaa acttctgaaa ggcataacaa ataatctgat
     1381 tgttgttttg agctgaattt ttacagacag actctggaca caaagattta tcctaaacct
     1441 tgaaagattg ataaaataaa agaacataga ctgataacac accaactaag actgcttacc
     1501 tctccaaaat gtcgatgccc attcagtgtg tgccagctca agtaagccca gtaatatgtt
     1561 gtccggctga gggcttgttc atggggattg actttgtatg gtggttatcc tggcacagtt
     1621 ttctcagccg cgttcattgt cagtttctgt aaagactaaa acctccaaaa tacacacggg
     1681 cacatcacct cacgggcatc ctaatctccg ttggcattaa atctagccac tggaatgcca
     1741 agcacctaca aacttaactc aatcctggca ggttcgccgg accagacagc gcagattcca
     1801 gtgaacacat gcgggaggca ctatcatcac aattaagggc tcacgctgta atccagccgg
     1861 tatctgcgag aactgaaatc cgatcagctc tttggataaa tactgtacag agagtgatgg
     1921 agggaagagg ggaaggggag aacacagtgg gagggctgca gtgaaagagg caaatgaacg
     1981 tcattcacat aattattcca aaaaaggtcc gtagtgggct cgtctgccag gggccgtcag
     2041 tatcactcta tttaagtgcg tagacagaag atttgcagaa gaggcccaga ctcaaaggat
     2101 ccaccggtcg ccaccatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc
     2161 ctgatcgagc tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag
     2221 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct
     2281 gtgccctggc ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac
     2341 cccgatcaca tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag
     2401 gagcgcacca tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc
     2461 gagggcgata ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc
     2521 aacatcctgg gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc
     2581 gacaaggcca agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc
     2641 agcgtgcagc tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg
     2701 ctgcccgata accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag
     2761 cgcgatcaca tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat
     2821 gagctgtaca agtgagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2881 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2941 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3001 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3061 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3121 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3181 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3241 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3301 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3361 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3421 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3481 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3541 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3601 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3661 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3781 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3901 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3961 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4021 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4081 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4141 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4201 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4261 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4321 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4381 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4441 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4501 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4561 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4621 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4681 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4741 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4801 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4861 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5041 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5101 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5161 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5221 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5281 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
