LOCUS Exported 5137 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION aA-crystallin promoter fragment in GFP plasmid to assess activity in larval embryos. ACCESSION . VERSION . KEYWORDS Danio alpha A-crystallin 1kb promoter/AcGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5137) 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 5137) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5137 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 1078..1087 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1084..1803 /codon_start=1 /product="Aequorea coerulescens GFP" /label=AcGFP1 /note="mammalian codon-optimized" /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF GFVTAAAITHGMDELYK" polyA_signal 1925..2046 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1962..1981) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2016..2035 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2053..2508) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2190..2211) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2402..2421 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2535..2639 /gene="bla" /label=AmpR promoter promoter 2641..2998 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2664..2684) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2849..2984 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2911..2930 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3033..3827 /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(3087..3106) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3697..3716 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4015..4034) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4059..4106 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4435..5023 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4924..4943 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattac tagcgctacc ggactcagat ctcgagtgga gacccctgat taataaaggg 61 acttagctga aaataaaata aatgaatgaa tgttttgtga taataaattt gatttgggca 121 gatctcctac ttagcttaat cacatgcatg gaatatattt atttagtgtg gtaagctaat 181 taacataatg acttcaaaac agcagttttg aataacagtt cagtaagtat ggcaaaacaa 241 taaatcttat tactcctgat ttgtccttaa attgtcattt ttaatatggc aataaacgat 301 cttcagagaa acttgcattg gtaaatagtt tcagcctatg catatgtagt tattcaaact 361 gtaatgacta tcttacagta ttatatgttt ataatagttt gatccaataa atccagattt 421 atatgctttt caacataaaa ttgtgaaatg cacagaatgt acatatacaa tacatttcct 481 acaataatgc tctcttttct attaatagtg tgcattcagt gcaggatgtt atgtatttat 541 tttctgtagt gcaaactttt tcaaagaatt ttccagtcta agcttttcag tcaagaaaaa 601 aaaaagcgat tgattatctg tgatatttaa aaaaattagc tcttaaatct ggtgaaccaa 661 gtggccacaa cacattaaat actgacgttc tacagccatg tagcttcatc tggtctggtt 721 tgttttggca ggcacttatt ttccggtgat agttggccac aagagtattg tctgagccat 781 tcagtgctag actgtcattc tcaggtcaga gtccaccccg ctgattctgc tgacaacact 841 gttcccacca tgagataatg ccattccaga gagatccatt tgtaagcccc tctttctgca 901 gcacaggtat ataaccaggg gtctgcctcc actaaggccg gcacacatca tttggggatc 961 tttgtactgt actagcctct attagcctcc tacttgcgtt cagttcacat cttttgttca 1021 tcaggtgcaa taagttacag taaggagtta ccaggtctga cattggatcc accggtcgcc 1081 accatggtga gcaagggcgc cgagctgttc accggcatcg tgcccatcct gatcgagctg 1141 aatggcgatg tgaatggcca caagttcagc gtgagcggcg agggcgaggg cgatgccacc 1201 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcctgt gccctggccc 1261 accctggtga ccaccctgag ctacggcgtg cagtgcttct cacgctaccc cgatcacatg 1321 aagcagcacg acttcttcaa gagcgccatg cctgagggct acatccagga gcgcaccatc 1381 ttcttcgagg atgacggcaa ctacaagtcg cgcgccgagg tgaagttcga gggcgatacc 1441 ctggtgaatc gcatcgagct gaccggcacc gatttcaagg aggatggcaa catcctgggc 1501 aataagatgg agtacaacta caacgcccac aatgtgtaca tcatgaccga caaggccaag 1561 aatggcatca aggtgaactt caagatccgc cacaacatcg aggatggcag cgtgcagctg 1621 gccgaccact accagcagaa tacccccatc ggcgatggcc ctgtgctgct gcccgataac 1681 cactacctgt ccacccagag cgccctgtcc aaggacccca acgagaagcg cgatcacatg 1741 atctacttcg gcttcgtgac cgccgccgcc atcacccacg gcatggatga gctgtacaag 1801 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc 1861 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt 1921 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 1981 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 2041 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt 2101 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag 2161 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga 2221 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg 2281 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc 2341 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg 2401 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc 2461 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt 2521 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt 2581 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc 2641 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 2701 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 2761 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 2821 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 2881 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 2941 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga 3001 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3061 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3121 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3181 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc 3241 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 3301 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 3361 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 3421 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 3481 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 3541 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg 3601 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 3661 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 3721 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 3781 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 3841 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 3901 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 3961 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac 4021 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa 4081 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc 4141 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc 4201 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg 4261 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta 4321 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg 4381 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga 4441 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt 4501 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag 4561 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa 4621 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag 4681 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca 4741 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac 4801 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa 4861 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc 4921 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg 4981 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc 5041 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc 5101 ccctgattct gtggataacc gtattaccgc catgcat //