LOCUS Exported 6729 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mRNA synthesis of tol1 transposase codon modified for zebrafish. ACCESSION . VERSION . KEYWORDS pCS2-Tol1.zf1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6729) AUTHORS Horstick EJ, Jordan DC, Bergeron SA, Tabor KM, Serpe M, Feldman B, Burgess HA TITLE Increased functional protein expression using nucleotide sequence features enriched in highly expressed genes in zebrafish. JOURNAL Nucleic Acids Res. 2015 Jan 27. pii: gkv035. PUBMED 25628360 REFERENCE 2 (bases 1 to 6729) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6729 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" promoter complement(2756..2773) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2778..2912 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2802..2821) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2856..2875 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3025..3045) /label=T3 /note="T3 promoter, forward primer" promoter complement(3025..3043) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3064..3080) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3064..3080) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3077..3099) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3088..3104 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(3112..3142) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3157..3178 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3295..3312) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3466..4054) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3546..3565) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4225..5085) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 4848..4867 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5086..5190) /gene="bla" /label=AmpR promoter rep_origin complement(5216..5671) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5353..5374) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 5565..5584 /label=F1ori-R /note="F1 origin, reverse primer" promoter 5745..6729 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atcccatcga ttcgaattcg agctcggtac ctcgcgaatg 121 catctagata cccaatgcca tcatggaaaa aaaacgatca aaaccctcag gcgcgcaatt 181 cagaaagaag agaaaggagg aggaggagaa gagagacaag gagaagggcg ctctgctgag 241 atactttgga tctagcacca ctgctcagga tgagaccagc acctctctgc ctgccatctc 301 ttctgccact gtgactgtgt ctcctcctca ggatgagctg cctagcacct cttctgccac 361 ccatgtggtg cctcagctgc tgcctgagca gtcttttgac tctgaggctg aggatgtggt 421 gcctagcacc agcacccagc tggagacctc tgagatgcct ggagatgaga cccctctgac 481 cccaactgct gaggaccagc ctctgccaac tgaccctgcc aagtggcctt ctcctctgac 541 tgacagaatc agaatggagc tggtgagaag aggaccttct agcatccctc ctgactttgt 601 gttcccaaga aatgactctg atggacgctc ttgccaccac cactacttca gaaagaccct 661 ggtgtctgga gagaagattg ccagaacctg gctgatgtac agcaaggtga agaactctct 721 gttctgcttc tgctgcaagc tgttcagcaa caagaacatc aacctgacca cctctggaac 781 tgccaactgg aagcatgcta gcacctacct gactgctcat gagaagtctc ctgagcacct 841 gaactgcatg aaggcttgga aggagctgtc tggaagaatc cgctctggaa agaccattga 901 caagcaggag atggctctgc tggaggagga gagagtgaga tggagagctg tgctgaccag 961 actgattgcc attgtgcagt ctctggctgt gagaaacctg gctctgagag gacacactga 1021 gaccctgttc acctctagca atggaaactt cctgaaggag gtggagctga tggccagatt 1081 tgacccaatc atgaaggacc acctgaacag agtgctgaga ggaactgctt ctcacaactc 1141 ttacattgga caccatgtgc agaatgagct gattgacctg ctgtctagca agatcctgtc 1201 tgccattgtg gatgacatca agaaggccaa gtacttcagc atcatcctgg actgcaccct 1261 ggacatctct cacactgagc agctgtctgt gatcatcaga gtggtgtctc tgatggagaa 1321 gcctcagatc agagagcact tcatgggatt cctggaggct gaggagagca ctggacagca 1381 cctggctagc atgatcctga acagactgga ggagctggga atctcttttg aggattgcag 1441 aggacagtct tatgacaatg gagccaacat gaagggaaag aacaagggag tgcaggccag 1501 actgctggag aagaacccac gcgctctgtt cctgccttgt ggcgctcaca ccctgaacct 1561 ggtggtgtgt gatgctgcca agcgctctgt ggatgccatg tcttactttg gagtgctcca 1621 gaagctgtac accctgttct ctgcttctgc tcagagatgg gccatcctga agtctcaggt 1681 gagcatcacc ctgaagtctt ggactgagac cagatgggag agcaagatca agagcattga 1741 gccaatgaga taccagggag ctgctgtgag agaggctctg attgaggtga gagacaagac 1801 caaggaccct gtgatcaagg ctgaggctca gtctctgtct gaggaggtgg gatcttacag 1861 attcaacatc tgcactgtgg tgtggcatga catcctgagc accatcaagc atgtgagcaa 1921 gctgatgcag tctccaaaca tgcatgtgga cctggctgtg tctctgctga agaagactga 1981 gcagtctctc cagtcttaca gagccaatgg atttgtgaat gctcagatgg ctgccaagga 2041 gatgtgcaag gagatgaatg tggaggccat cctgaagcag aagagaatca gaagcaccaa 2101 gtgccagttc tcttatgagt ctcatgatga gcctttctct gatgctctga agaagctgga 2161 ggtggagttc ttcaatgtgg tggtggatga ggctctgtct gccattgctg agagattcag 2221 caccctggag gtggtgcaga acagatttgg agtgctgacc aacttccctt ctctgggaga 2281 tgaggagctg actgagcagt gtgaggctct gggaaacatc ctgcactttg agaagaactg 2341 ggacctggac agcagagagc tggtgcagga gatcaagaac ctgccaaacc tgcctagcac 2401 caccccttct ctgctggagc tgatctcttt catgtctgac aaggacctgt ctgaaatcta 2461 cccaaacttc tggactgctc tgagaattgc tctgaccctg cctgtgactg tggctcaggc 2521 tgagcgctct ttcagcaagc tgaagctgat caagtcttac ctgagaagca ccatgtctca 2581 ggagagactg accaacctgg ctgtggtgag catcaaccac tctgtgggag agcaaatctc 2641 ttatgatgat gtgattgatg agtttgctag cagaaaggcc agaaaggtga gattctaaaa 2701 ttgggtcatc ggatcccggg cccgtcgact gcagaggcct ctcgagcctc tagaactata 2761 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca 2821 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat 2881 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt 2941 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt 3001 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc 3061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 3121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 3181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 3241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 3301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 3361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 3421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 3481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 3541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 3601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 3661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 3721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 3781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 3841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 3901 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 3961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 4021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 4081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 4141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 4201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 4261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 4321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 4381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 4441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 4501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 4561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 4621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 4681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 4741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 4801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 4861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 4921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 4981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 5041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 5101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 5161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt 5221 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt 5281 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg 5341 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc 5401 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca 5461 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga 5521 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa 5581 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc 5641 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt 5701 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca 5761 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca 5821 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca 5881 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg 5941 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc 6001 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc 6061 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata 6121 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat 6181 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag 6241 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg 6301 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg 6361 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc 6421 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc 6481 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag 6541 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat 6601 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt 6661 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt 6721 ttagggaac //