LOCUS Exported 7587 bp ds-DNA circular SYN 25-APR-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7587) AUTHORS Karl Clark TITLE Direct Submission JOURNAL Exported Thursday, Jul 8, 2021 from SnapGene 5.0.4 https://www.snapgene.com FEATURES Location/Qualifiers source 1..7587 /organism="synthetic DNA construct" /mol_type="other DNA" source 1049..4525 /organism="recombinant plasmid" /mol_type="other DNA" /note="/locus_tag=source:recombinant plasmid" /note="color: #ffc0cb" source 1193..1249 /organism="recombinant plasmid" /mol_type="other DNA" /note="/locus_tag=source:recombinant plasmid" /note="color: #ffc0cb" rep_origin complement(1..253) /direction=LEFT /locus_tag="ColE1 Ori" /label=ColE1 Ori /note="ColE1 Ori" /note="color: #ffc0cb" primer_bind 241..266 /label=Ori_F1 /note="color: black; sequence: ggaagcggaacagatttaaatggtac; added: 2021-03-01" misc_feature complement(267..457) /locus_tag="ITR(R)" /label=ITR(R) /note="ITR(R)" /note="Tol2 ITR(R)" /note="color: #80ffff; direction: LEFT" misc_feature 487..506 /locus_tag="5' Universal Guide" /label=5' Universal Guide /note="color: #00ffff" misc_feature complement(506..509) /locus_tag="BfuAI site 1 overhang" /label=BfuAI site 1 overhang /note="color: #00ff00; direction: LEFT" misc_feature complement(514..519) /locus_tag="BfuA site 1" /label=BfuA site 1 /note="color: #00ff00; direction: LEFT" promoter 520..641 /locus_tag="lacZ\promoter" /label=lacZ promoter /note="/vntifkey=30" /note="color: #ffc0cb" misc_feature 642..917 /locus_tag="lacZ'" /label=lacZ' /note="/vntifkey=21" /note="color: #ffc0cb" misc_feature 918..923 /locus_tag="BfuA site 2" /label=BfuA site 2 /note="color: #00ffff" misc_feature complement(928..938) /locus_tag="TAL3" /label=TAL3 /note="TAL3" /note="color: #ffc0cb; direction: LEFT" misc_feature 928..931 /locus_tag="overhang5'3'" /label=BfuAI site 2 overhang 5'3' /note="color: #00ffff" misc_feature 940..971 /locus_tag="rox(1)" /label=rox /note="color: #ff0080; direction: RIGHT" misc_feature 977..1010 /locus_tag="loxp" /label=loxp /note="color: #80ff80; direction: RIGHT" misc_feature 1015..1048 /locus_tag="lox2272" /label=lox2272 /note="color: #8080ff; direction: RIGHT" intron 1060..1192 /locus_tag="BP/SA" /label=BP/SA /note="BP/SA" /note="BP/SA from carp beta actin intron 1" /note="color: #ffc0cb" primer_bind complement(1168..1202) /label=2A_SA_R0 /note="color: black; sequence: agttcgtggcctgtaaatgaatgagaaaaccggtt; added: 2021-03-01" primer_bind 1183..1210 /label=SA_2A_F0 /note="color: black; sequence: tcatttacaggccacgaacttctctctg; added: 2021-03-01" CDS 1193..1249 /codon_start=1 /locus_tag="2A peptide from porcine teschovirus-1 polyprotein" /product="2A peptide from porcine teschovirus-1 polyprotein" /label=2A peptide from porcine teschovirus-1 polyprotein /label=2A peptide from porcine teschovirus-1 polyprotein /note="P2A" /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /note="color: #00ff80" /translation="ATNFSLLKQAGDVEENPGP" primer_bind complement(1232..1258) /label=RFP_2A_R /note="color: black; sequence: ggaggaggcaggaccggggttttcttc; added: 2021-01-08" primer_bind 1241..1267 /label=2A_RFP_F /note="color: black; sequence: cccggtcctgcctcctccgaggacgtc; added: 2021-01-08" CDS 1250..1924 /locus_tag="mRFP*" /label=mRFP* /note="mRFP*" /note="color: #ffc0cb; direction: RIGHT" primer_bind complement(1565..1602) /label=mRFP_R /note="color: black; sequence: atgaactcgccgtcctgcagggaggagtcctgggtcac; added: 2021-03-01" polyA_signal 1933..3108 /locus_tag="ocean pout terminator" /label=ocean pout terminator /note="ocean pout terminator" /note="3'UTR, poly(A) signal, and putative boundary element from ocean pout (FRM- Pat Gibbs)." /note="color: #ffc0cb" misc_feature 3152..3512 /locus_tag="gamma-crystallin" /label=gamma-crystallin /note="gamma-crystallin" /note="Xenopus" /note="color: #ffc0cb" intron 3565..3762 /locus_tag="mIntron" /label=mIntron /note="mIntron" /note="carp bactin shortened intron I" /note="color: #ffc0cb" primer_bind complement(3733..3757) /label=2A_SA_R0 /note="color: black; sequence: agttcgtggcctgtaaatgaatgagaaaaccggtt; added: 2021-03-01" misc_signal 3783..3812 /locus_tag="NLS" /label=NLS /note="NLS" /note="color: #ffc0cb" CDS 3813..4514 /codon_start=1 /locus_tag="monomeric blue fluorescent protein" /product="monomeric blue fluorescent protein" /label=monomeric blue fluorescent protein /note="TagBFP" /note="mammalian codon-optimized" /note="color: #ffc0cb" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" misc_feature 4526..5124 /locus_tag="Danio b-Actin 3' UTR" /label=Danio b-Actin 3' UTR /note="color: #0000ff" misc_feature 5105..5529 /locus_tag="Danio b-Actin poly(A) putative" /label=Danio b-Actin poly(A) putative /note="color: #004080" misc_feature complement(5536..5569) /locus_tag="loxp(1)" /label=loxp /note="color: #00ff80; direction: LEFT" misc_feature complement(5575..5608) /locus_tag="lox2272(1)" /label=lox2272 /note="color: #800080; direction: LEFT" misc_feature complement(5615..5646) /locus_tag="rox" /label=rox /note="color: #ff00ff; direction: LEFT" misc_feature 5656..5658 /locus_tag="TAL3(1)" /label=TAL3 /note="TAL3" /note="color: #ffc0cb" misc_feature 5656..5658 /locus_tag="overhange3'5'" /label=BspQI site 1 overhang /note="color: #00ffff" misc_feature 5660..5666 /locus_tag="BspQ1 site1" /label=BspQ1 site1 /note="color: #00ffff" intron 5675..5763 /locus_tag="Carp beta-actin intron 1 filler" /label=Carp beta-actin intron 1 filler /note="Carp beta-actin intron 1 filler" /note="color: #ffc0cb" misc_feature 5766..5772 /locus_tag="BspQ1site2" /label=BspQ1site2 /note="color: #00ffff" misc_feature 5774..5776 /locus_tag="BspQI site 2 overhang" /label=BspQI site 2 overhang /note="color: #00ffff" misc_feature complement(5777..5796) /locus_tag="Universal Guide 3'" /label=Universal Guide 3' /note="color: #00ff00; direction: LEFT" misc_feature 5838..6052 /locus_tag="ITR(L)" /label=ITR(L) /note="ITR(L)" /note="Tol2 ITR(L)" /note="color: #80ffff" CDS 6117..6926 /locus_tag="Kan(R)" /label=Kan(R) /note="Kan(R)" /note="color: #ffc0cb; direction: RIGHT" rep_origin complement(6959..7587) /direction=LEFT /locus_tag="ColE1 Ori(1)" /label=ColE1 Ori /note="ColE1 Ori" /note="color: #ffc0cb" ORIGIN 1 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 61 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 121 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 181 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 241 ggaagcggaa cagatttaaa tggtaccgag cagaggtgta aaaagtactc aaaaatttta 301 ctcaagtgaa agtacaagta cttagggaaa attttactca attaaaagta aaagtatctg 361 gctagaatct tacttgagta aaagtaaaaa agtactccat taaaattgta cttgagtatt 421 aaggaagtaa aagtaaaagc aagaaagaaa actagagtgg tctcccttta gtgagggtca 481 attgatggga ggcgttcggg ccacagcgga cacgcaggtg cgcaacgcaa ttaatgtgag 541 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg 601 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa 661 gcgcgcaatt aaccctcact aaagggaaca aaagctggag gacctcgagg gggggcccgg 721 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac 781 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 841 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca 901 gcctgaatgg cgaatggacc tgccacactt ccggccgcat aactttaaat aatgccaatt 961 atttaaagtt actatcataa cttcgtatag catacattat acgaagttat caccataact 1021 tcgtatagga tactttatac gaagttatcc taggcttacc acccactagc ggtcagactg 1081 cagattgcag cacgaaacag gaagctgact ccacatggtc acatgctcac tgaagtgttg 1141 acttccctga cagctgtgca ctttctaaac cggttttctc attcatttac aggccacgaa 1201 cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccggtcctg cctcctccga 1261 ggacgtcatc aaggagttca tgcgcttcaa ggtgcgcatg gagggctccg tgaacggcca 1321 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa 1381 gctgaaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt 1441 ccagtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct 1501 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 1561 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg 1621 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc 1681 ctccaccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agatgaggct 1741 gaagctgaag gacggcggcc actacgacgc cgaggtcaag accacctaca tggccaagaa 1801 gcccgtgcag ctgcccggcg cctacaagac cgacatcaag ctggacatca cctcccacaa 1861 cgaggactac accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggagc 1921 ctaagactag taattaagtc tcagccaccg ttaactgaac atgtcaaaac ctgtggagac 1981 tgttgagatt tgatgttctg aaaagataaa gcctataaat aaaatgttgc ccaaatttcc 2041 tgcctgatgt ttttctttgt ctttgctaca tggctttgct gctcggatcg gctcactctg 2101 tgtatgccac gttcactttg tactctcctt ctcacggtag gtttattatt tttagatgtg 2161 cagttagttt ctgtgaaata acacaccaca cactgatatt gtctgtgcat tgacttggtg 2221 agtgcacatt gtttttgatc ttgacatatt tatatttgat tgatcaggtg aactgtgtga 2281 atctaaagtg ctccatacag atgttctgca ttgaaaatat tctcatttta ttagtggaag 2341 tgagtgtatg ccacatccaa tcaatttcag caaacacccc agtatgattt aatgcaaaaa 2401 aatgaaggta tcaaacacgc attactactt tgcagttaaa tatttaacat ttattccaac 2461 acgaaaaaaa gcagtaaata acactttgac aaacacgtca ggacatctta tttttgtcac 2521 cctcacaggc aatttagtat aatatattat atatatatat atatcatata ataatattca 2581 gtataatata tatatatata tcatattata atattcagta taatataaaa cacaaacaca 2641 tatatgtata atataatata acatttttat ttattgagat gcctctatgg accgtgttat 2701 aagaagtaaa gatcaggaga agtaaacatg aagtgtaatt atgaatactg atgttaaatt 2761 aagctatgat gagttttcac tgttaattta ccatctcaat taaatgttga tgcctccatg 2821 accaagttaa gcagatgaga ctgagacaac tgtagaagac aagatgttca ctttgctgaa 2881 tatagctggc ttgacagtta tctatgactc tataaatata tatatatttt tttttttata 2941 aaatgattta tttataacta tatatccatt tctcagacag gtgcttcata tccctcactc 3001 ccgtagctgt ccatgctgga tctgtccccg ttgtttttaa aaagctaaat aagttattaa 3061 catgactgca tccagcgagc caaacctgtc tggtgtacag ctaccagaga agcttgagat 3121 cctagtcacc ggcgttccta gttctttaaa ctgttacaaa gtgtttggac cagcaactga 3181 attgcactgt aaattaccac atcagggtct ccggggtcaa cgctgctaag caatggactg 3241 gaaaatttgt gacattgagt ttttggaata actgggaatg ctggataact ctttgggtaa 3301 agctatggaa caaaagtacc agcatgaact aagaaatgac tgtcacagca caacatcaaa 3361 cacaatgcaa gacaaaggtt ctctgctttt gccttacttg acagaaaaag taagctctag 3421 gttcatcgca tgcagacagc agcaatacat gacatatata aaggctggct cttttctccc 3481 ttccattgaa ctgaaacttc cactcagtca gacttgcggg ggatccacag cctgactaac 3541 tcgacaaacc cccccaaacc taaggtgagt tgatctttaa gctttttaca ttttcagctc 3601 gcatatatca attcgcgcgc attgtcacac tagcacccac tagcggtcag actgcagaca 3661 aacaggaagc tgactccaca tggtcacatg ctcactgaag tgttgacttc cctgacagct 3721 gtgcactttc taaaccggtt ttctcattca tttacagttc agccggaatt cacctcgaga 3781 ccatggctcc aaagaagaag cgtaaggtaa ccatgagcga gctgattaag gagaacatgc 3841 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg 3901 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc 3961 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca 4021 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat 4081 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc 4141 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg 4201 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg 4261 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc 4321 tgatcgcaaa catcaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc 4381 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct 4441 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc 4501 acaagctcaa ttaaagctga gatctacgga ctgttaccac ttcacgccga ctcaaactgc 4561 gcagagaaac ttcaaacgac aacattggca tggcttttgt tatttttggc gcttgactca 4621 ggatctaaaa actggaacgg tgaaggtgac ggcaatgttt tttggcaaat aagcatcccc 4681 gaagttctac aatgcatctg aggactccat gttttgtttt ctttagtcat tccaaatgtt 4741 tgttaaatgc attgttccga aacttatttg cctctatgaa ggctgcccag taattgggag 4801 catacttaac attgtagtat tgtatgtaaa ttatgtaaca aaacaatgtc tgggtttttg 4861 tacttttcag ccttaaaatc ttgggtttat ttttttttct ttgttccaaa aaaacgaagc 4921 tttaccattc aagatgtaaa ggtttccatt ccccctgggc atattgtaaa agctgtgtgg 4981 aacgtggcgg tgccagacat ttggtggggc caacctgtac actgactaat tcaattccaa 5041 taaaagtgca catgtaagac atcctacccc tgtctcattt cctgtcttgg ctctgagtgg 5101 cttcaataaa aatggctttt gtgaagttgc accatgttaa ttacatttca gcactttcac 5161 accaggtttt tgactaattt gaatttatcg aagtttgacc tttttttttt tttttcttca 5221 agctaatttg aggttatcaa accatttcat tggaggaaac ttaatgttta aaagcataaa 5281 ttcgaaacat gattcattta tttaactgct taatggtgaa acaaacttca agtaatacac 5341 caagtgttcc ctcatatgaa ggtagctaac tgcaaacggc cttaactttc catcttaagt 5401 tgagctacat aaaggcttta acaggaagga actgccagtt taaaagtcaa accaccatga 5461 ctgatataaa ttcttgtcca tataaccttg cgtttcacac aatatagcta tctactgcta 5521 aataaattag agctcataac ttcgtataat gtatgctata cgaagttatg ttagataact 5581 tcgtataaag tatcctatac gaagttatgc gagctaactt taaataattg gcattattta 5641 aagttatgaa tcgataagtg aagagctatc cggatatata tgtatgttat cataactgta 5701 ctggctatgt caggtggtaa tgactgtaac gttacgttac tcgttgtagg cacgacattg 5761 aatatgctct tctccgctgt ggcccgaacg cctcccgcta gcgatgggat ctaatacgac 5821 tcactatagg gagaccaagg gaaaatagaa tgaagtgatc tccaaaaaat aagtactttt 5881 tgactgtaaa taaaattgta aggagtaaaa agtacttttt tttctaaaaa aatgtaatta 5941 agtaaaagta aaagtattga tttttaattg tactcaagta aagtaaaaat ccccaaaaat 6001 aatacttaag tacagtaatc aagtaaaatt actcaagtac tttacacctc tgggatccag 6061 ctgcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga 6121 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat 6181 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt 6241 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg 6301 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca 6361 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa 6421 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc 6481 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg 6541 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga 6601 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata 6661 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc 6721 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag 6781 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac 6841 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc 6901 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaaa 6961 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 7021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 7081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 7141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 7201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 7261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 7321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 7381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 7441 gagataccta cagcgtgagc tatgagaaag cgccacggtt cccgaaggga gaaaggcgga 7501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 7561 aaacgcctgg tatctttata gtcctgt //