LOCUS Exported File 6424 bp ds-DNA circular SYN 29-SEP-2014 DEFINITION . ACCESSION . VERSION . KEYWORDS pSBbi_RB SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6424) AUTHORS Eric Kowarz TITLE . JOURNAL . FEATURES Location/Qualifiers source 1..6424 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 8..11 /note="tata" /note="color: #cc0000" misc_feature 8..11 /note="tata" /note="color: #cc0000" misc_feature 12..236 /note="5'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 242..283 /note="5'-proxITR" /note="modified region proximal to 5'-ITR - contains CTCF binding sites" /note="color: #cc0000" polyA_signal complement(294..591) /note="bGH-pA" /note="bovine growth hormone polyadenylation signal" /note="color: #cc0000; direction: LEFT" misc_feature 592..604 /note="3'-SfiI" /note="GGCCTGTCAGGCC" /note="color: #a6acb3" misc_feature 605..800 /note="filler DNA" /note="color: #cc0000; direction: RIGHT" misc_feature 801..813 /note="5'-SfiI" /note="GGCCTCTGAGGCC" /note="color: #a6acb3" promoter complement(819..2011) /note="EF1a promoter" /note="constitutive promoter" /note="color: #cc0000; direction: LEFT" exon complement(819..837) /note="EF1a Exon2" /note="color: #cc0000; direction: LEFT" intron complement(838..1780) /note="EF1a Intron1" /note="color: #cc0000; direction: LEFT" promoter 2018..2629 /note="RPBSA promoter" /note="constitutive promoter" /note="color: #cc0000; direction: RIGHT" promoter 2018..2629 /note="RPBSA" /note="constitutive promoter" /note="color: #cc0000; direction: RIGHT" promoter 2018..2452 /note="RPL13a Promoter" /note="color: #cc0000; direction: RIGHT" exon 2452..2504 /note="RPL41 exon1" /note="color: #cc0000; direction: RIGHT" intron 2505..2619 /note="RPL41 intron1" /note="color: #cc0000; direction: RIGHT" exon 2620..2629 /note="RPL41 exon2" /note="color: #cc0000; direction: RIGHT" CDS 2643..3344 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein" /note="dTomato" /note="mammalian codon-optimized" /note="color: #ff0000" /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLYGMDELYK" CDS 3345..3410 /codon_start=1 /note="P2A" /note="color: #cc0000" /translation="GSGATNFSLLKQAGDVEENPGP" CDS 3411..3809 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /note="BSD" /note="confers resistance to blasticidin" /note="color: #ccffcc" /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 3827..4127 /note="rBG-pA" /note="rabbit beta-globin polyadenylation signal" /note="color: #cc0000; direction: RIGHT" misc_feature 4138..4179 /note="3'-proxITR" /note="modified region proximal to 3'-ITR - contains CTCF binding sites" /note="color: #cc0000" misc_feature 4180..4404 /note="3'-ITR" /note="Sleeping Beauty inverted tandem repeat" /note="color: #cc00cc" misc_feature 4405..4408 /note="tata" /note="color: #cc0000" rep_origin complement(4592..5180) /direction=LEFT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(5351..6211) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments:    1: 5351 .. 6142 / #ccffcc    2: 6143 .. 6211 / #ccffcc / signal sequence" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6212..6316) /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: LEFT" ORIGIN 1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact 61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta 121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta 181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt 241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca 301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt 361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca 421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc 481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga 541 aggcacagtc gaggctgatc agcgagctct agagaattga tccccaagct tggcctgaca 601 ggccttcgaa ttagagagaa ggaaagacgc aaccaggaag cccaccaaga gaaagaggca 661 tttcctgaaa agattcccct ttttggagag ccctacaagg tatttactga acactagaca 721 ttgaagtcct gatttatcac aatgttgaac cctatgatga aacaattcag tataattgag 781 ttcgaaactc gagccatggt ggcctcagag gccagcttgg ggtagttttc acgacacctt 841 aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc aagatccaaa 901 ctcaaaaagg gcaaattcca aggagaatta catcaagtgc caagctggcc taacttcagt 961 ctccacccac tcagtatggg gaaactccat cgcataaaac ccctcccccc aacctaaaga 1021 cgacgtactc caaaagctcg agaactaatc gaggtgcctg gacggcgccc ggtactccgt 1081 ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc 1141 acccgcccgc tctcccgagc gccgcgtcct ccattttgag ctccctgcag cagggccggg 1201 aagcggccat ctttccgctc acgcaactgg tgccgaccgg gccagccttg ccgcccaggg 1261 cggggcgata cacggcggcg cgaggccagg caccagagca ggccggccag cttgagacta 1321 cccccgtccg attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg 1381 ggacgcacgg gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc 1441 ttggcccgca tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa 1501 attttaaatg gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat 1561 tcgcacgcgg cggccccagc gcccaggcca agcctcaact caagcacgag gcgaaggggc 1621 tccttaagcg caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag 1681 aatcacgtac tgcagccagg ggcgtggaag taattcaagg cacgcaaggg ccataacccg 1741 taaagaggcc aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc 1801 cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct 1861 cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc 1921 tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg 1981 tgcgctctgc ccactgacgg gcaccggagc ctctaggcga gaccctgtct cacaaaataa 2041 agtaagcccg gactgagtgc ggaaaggcgg gcctggcggg tctggtctcc ccatgcgggc 2101 caccagaggc cctgcagcct tcagtcgctt gaaggggtaa tggcgcttcc actcacaaac 2161 atggcggaca gagcgtgtga acgagatgaa cagcccctca aaaatatggc cgccgaggct 2221 ggacggccgt gccccagcag caccgcctcc gcgccccacg tgatctctcg ccgggcacag 2281 cgctgaccgc ggaggtccaa ccggaagaat gtccggattg gacattcgga agagggcccg 2341 ccttccctgg ggaatctctg cgcacgcgca gaacgcttcg accaatgaaa acacaggaag 2401 ccgtccgcgc aaccgcgttg cgtcacttct gccgcccctg tttcaaggta tatagccgta 2461 gacggaactt cgcctttctc tcggccttag cgccattttt ttgggtgagt gttttttggt 2521 tcctgcgttg ggattccgtg tacaatccat agacatctga cctcggcact tagcatcatc 2581 acagcaaact aactgtagcc tttctctctt tccctgtaga aacctctgca cctgaggcca 2641 ccatgttgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg 2701 agggctccat gaacggccat gagttcgaga tcgagggcga gggcgagggc cgcccctacg 2761 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg 2821 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca 2881 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact 2941 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga 3001 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga 3061 agacaatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg 3121 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga 3181 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc 3241 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg 3301 gccgccacca cctgttcctg tacggcatgg acgagctgta caagggcagt ggagctacta 3361 acttcagcct gctgaagcag gctggtgacg tcgaggagaa tcctggcccc atggccatgc 3421 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc 3481 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca 3541 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca 3601 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca 3661 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga 3721 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc 3781 tgccctctgg ttatgtgtgg gagggctaat tcgaaggcct gtcgtgaatt cactcctcag 3841 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac 3901 cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat 3961 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg 4021 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat 4081 ttggtttaga gtttggcaac atatgccata tgctggctgc catgaactag ctactcggga 4141 ccccttaccg aaacatcgcc gcattctgca gaggagtcga gtgtatgtaa acttctgacc 4201 cactgggaat gtgatgaaag aaataaaagc tgaaatgaat cattctctct actattattc 4261 tgatatttca cattcttaaa ataaagtggt gatcctaact gacctaagac agggaatttt 4321 tactaggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg 4381 tatgtaaact tccgacttca actgtatagg gatccgcttc ctcgctcact gactcgctgc 4441 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4501 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4561 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4621 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4681 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4741 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4801 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4861 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4921 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4981 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 5041 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 5101 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 5161 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 5221 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 5281 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 5341 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 5401 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 5461 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 5521 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 5581 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 5641 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 5701 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 5761 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 5821 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 5881 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 5941 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 6001 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 6061 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 6121 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 6181 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 6241 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 6301 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg 6361 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa 6421 aatt //