LOCUS       sequence-193850-        4374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse aB crystallin 250bp promoter segment driving GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4374)
  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 4374)
  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 250bp promoter/AcGFP
FEATURES             Location/Qualifiers
     source          1..4374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      315..324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             321..1040
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    1162..1283
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1199..1218)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1253..1272
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1290..1745)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1427..1448)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1639..1658
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1772..1876
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1878..2235
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1878..2069
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1901..1921)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2086..2221
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2270..3064
                     /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             2270..3064
                     /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(2324..2343)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2934..2953
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3252..3271)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3296..3343
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3672..4260
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4161..4180
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttcggtg aaacaagacc
       61 atgacaagtc accggtcagc tcagccctgc ctgtgtttct cttttcttag ctcagtgagt
      121 accgggtatg tgtcaccctg ccaaatccct gatcacaagt ctccatgaac tggcggtgag
      181 ctgggataat aaaacccctg acctcaccat tccagaagct tcagaagact gcatatataa
      241 ggggccggct ggagctgctg ctgaaggagt tgaccagcca accgactctg cattcatcta
      301 gccacacacc ggtcgccacc atggtgagca agggcgccga gctgttcacc ggcatcgtgc
      361 ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg agcggcgagg
      421 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      481 tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag tgcttctcac
      541 gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct gagggctaca
      601 tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc gccgaggtga
      661 agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg
      721 atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat gtgtacatca
      781 tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
      841 atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc gatggccctg
      901 tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag gaccccaacg
      961 agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc acccacggca
     1021 tggatgagct gtacaagtga gcggccgcga ctctagatca taatcagcca taccacattt
     1081 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     1141 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     1201 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1261 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     1321 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     1381 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     1441 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     1501 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     1561 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     1621 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     1681 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     1741 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1861 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     1921 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     1981 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2041 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2101 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2161 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2221 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     2281 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     2341 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     2401 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     2461 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     2521 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     2581 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     2641 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     2701 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     2761 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     2821 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     2881 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     2941 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3001 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3061 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     3121 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3181 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     3241 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3301 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3361 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3421 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3481 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     3541 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3601 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3661 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3721 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3781 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     3841 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3901 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3961 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4021 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4081 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4141 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4201 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     4261 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     4321 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
