LOCUS       sequence-193852-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse aB crystallin 1.5kb promoter segment driving GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  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 5607)
  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: Mouse aB crystallin 1.5kb promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..5607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1548..1557
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1554..2273
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    2395..2516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2432..2451)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2486..2505
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2523..2978)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2660..2681)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2872..2891
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3005..3109
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3111..3468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3111..3302
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3134..3154)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3319..3454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3503..4297
                     /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             3503..4297
                     /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(3557..3576)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4167..4186
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4485..4504)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4529..4576
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4905..5493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5394..5413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagaaag caagaggcag gatgagggtg
       61 gatgcagtgg gcacagggat gaaaggcgat atctggagac atgggccaga agagcaggtg
      121 ggaactgctg aagaggggct gggaaaggaa aggagatagc catggaggaa gcgagctgga
      181 attgggagga cagacgcgaa atcagaaggt actctgaagc ctgcaaagcc cagttaccca
      241 aaaccctacc ggtctaaagt ttatcctgag gtctgtagca agaccagaga gagtcttgct
      301 tgacggggga atctggaatg ggagatggtt cacctcagga gttcacttaa gccccaggta
      361 gagtggaggc cagtaaagct gggcagtgag cccagaattt caggaggtgg gggtgggagg
      421 cagcatctga gtcctacctt ctccgaagcg ctgctcgccc agacggctgg ggttggcaaa
      481 ttcgtactcg gcagtggctg ggtgggcgtg tggcactgtg cggcccgaca tgactgctgt
      541 tgcgactagt agccctgatc ccaagtactg ccccaacaag atcaaggcga ccccaccggc
      601 ttcagggaga gccagggctc agtcagagtg ggggcggggt ctacaccacc caaaatagtg
      661 cagagcctct gggggtgggg gagggctggg agcctaagtc tagagtgcag ctatgagggt
      721 gtgatcagcc tggaccccct aagtcccctc ctctgcacac ccttcagctg tcgcaggggc
      781 ctggtaggac agctgagggt cctggtcctg gagtgagctc ctgctctgca ggagggcaag
      841 gagaggccta gttgggcctt caccagttgt acattccaca tcaccctttg tccttctcag
      901 tctcaggcac tgtgcacatt acttaaaaaa aaaaaagact taatgttcta tgagccacat
      961 agcatccaca atgcaagaaa cattttctgt ctttgtaagg tcagtgtctt ctgaacctag
     1021 atcagctcag ggttccagtc agacacctag ttctgctctc ctctaggact ccacaaagag
     1081 ttaatgtccc tggggctaag cctaggaaga ttccagtccc tgcccaggcc caagatagtt
     1141 gctggctcaa ttcccctggc atgcgagact ggagaggagg aggggcccac cagcagctgc
     1201 ttgggattcc aggctccgtc ctagctccag agaacaagga tggggtgggt gccactgggt
     1261 gtggacagag agctagtgaa acaagaccat gacaagtcac cggtcagctc agccctgcct
     1321 gtgtttctct tttcttagct cagtgagtac cgggtatgtg tcaccctgcc aaatccctga
     1381 tcacaagtct ccatgaactg gcggtgagct gggataataa aacccctgac ctcaccattc
     1441 cagaagcttc agaagactgc atatataagg ggccggctgg agctgctgct gaaggagttg
     1501 accagccaac cgactctgca ttcatctagc cacaggatcc accggtcgcc accatggtga
     1561 gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg aatggcgatg
     1621 tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc tacggcaagc
     1681 tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc accctggtga
     1741 ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg aagcagcacg
     1801 acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc ttcttcgagg
     1861 atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc ctggtgaatc
     1921 gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc aataagatgg
     1981 agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag aatggcatca
     2041 aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg gccgaccact
     2101 accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac cactacctgt
     2161 ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg atctacttcg
     2221 gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag tgagcggccg
     2281 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2341 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2401 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2461 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     2521 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     2581 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     2641 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     2701 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     2761 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     2821 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     2881 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     2941 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3001 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3061 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3121 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3181 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3241 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3301 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3361 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3421 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     3481 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3541 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3601 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3661 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     3721 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3781 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3841 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3901 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3961 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4021 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4081 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4141 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4201 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4261 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4321 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4381 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4441 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     4501 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     4561 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4621 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4681 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4741 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     4801 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     4861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5041 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5101 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5161 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5221 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5281 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     5341 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5401 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5461 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5521 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5581 gtggataacc gtattaccgc catgcat
//
