LOCUS Exported 6835 bp ds-DNA circular SYN 15-DEC-2022 DEFINITION SB-transposon with constitutive bi-directional promoter, one side: SfiI cloning site for GOI (contains filler DNA), other side: GFP and neomycin resistance gene. ACCESSION . VERSION . KEYWORDS pSBbi-GN SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6835) AUTHORS Kowarz E, Loescher D, Marschalek R TITLE Optimized Sleeping Beauty transposons rapidly generate stable transgenic cell lines. JOURNAL Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. PUBMED 25650551 REFERENCE 2 (bases 1 to 6835) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 15, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6835 /organism="synthetic DNA construct" /mol_type="other DNA" polyA_signal 324..548 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 537..554 /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" regulatory 794..803 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(829..2010) /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" primer_bind complement(861..881) /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 2643..3359 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2643..3359 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2688..2709) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2949..2968) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3296..3317 /label=EGFP-C /note="EGFP, forward primer" CDS 3369..3425 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 3426..4220 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3480..3499) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4090..4109 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4320..4339) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 4385..4440 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(4439..4458) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4832..4849) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5003..5591) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5083..5102) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5762..6622) /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 6385..6404 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6623..6727) /gene="bla" /label=AmpR promoter 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 ttagtgagaa 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 gcccaagcca ggcctcaact 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 gagctgttca ccggggtggt gcccatcctg gtcgagctgg 2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct 2761 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca 2821 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga 2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct 2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc 3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc 3061 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga 3121 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg 3181 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc 3241 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg 3301 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg 3361 gcagtggagc tactaacttc agcctgctga agcaggctgg tgacgtcgag gagaatcctg 3421 gccccatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 3481 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 3541 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 3601 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 3661 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 3721 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 3781 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 3841 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 3901 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc 3961 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 4021 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 4081 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 4141 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 4201 ttcttgacga gttcttctga ttcgaaggcc tgtcgtgaat tcactcctca ggtgcaggct 4261 gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat 4321 ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct 4381 ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca 4441 ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag 4501 agtttggcaa catatgccat atgctggctg ccatgaacta gctactcggg accccttacc 4561 gaaacatcgc cgcattctgc agaggagtcg agtgtatgta aacttctgac ccactgggaa 4621 tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc tactattatt ctgatatttc 4681 acattcttaa aataaagtgg tgatcctaac tgacctaaga cagggaattt ttactaggat 4741 taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt tggctaaggt gtatgtaaac 4801 ttccgacttc aactgtatag ggatccgctt cctcgctcac tgactcgctg cgctcggtcg 4861 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4921 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4981 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5041 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5101 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5161 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5221 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5281 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5341 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5401 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5461 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5521 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 5581 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 5641 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 5701 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 5761 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 5821 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 5881 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 5941 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 6001 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6061 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6121 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6181 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 6241 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 6301 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 6361 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 6421 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 6481 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 6541 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 6601 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 6661 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 6721 ttccgcgcac atttccccga aaagtgccac ctgatgcggt gtgaaatacc gcacagatgc 6781 gtaaggagaa aataccgcat caggaaattg taagcgttaa tattttgtta aaatt //