LOCUS       Exported                7051 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  SB-transposon with constitutive bi-directional promoter, one side: 
            SfiI cloning site for GOI (contains filler DNA), other side: RFP and
            hygromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    pSBbi-RH
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7051)
  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 7051)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7051
                     /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..3344
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2776..2794)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3044..3063
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3238..3261
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3354..3410
                     /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             3411..4436
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(4536..4555)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4601..4656
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4655..4674)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5048..5065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5219..5807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5299..5318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5978..6838)
                     /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     6601..6620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6839..6943)
                     /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 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 atgaaaaagc
     3421 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     3481 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     3541 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     3601 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     3661 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     3721 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     3781 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     3841 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     3901 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     3961 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     4021 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     4081 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     4141 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     4201 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     4261 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     4321 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     4381 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagttcg
     4441 aaggcctgtc gtgaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     4501 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     4561 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     4621 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     4681 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc
     4741 tggctgccat gaactagcta ctcgggaccc cttaccgaaa catcgccgca ttctgcagag
     4801 gagtcgagtg tatgtaaact tctgacccac tgggaatgtg atgaaagaaa taaaagctga
     4861 aatgaatcat tctctctact attattctga tatttcacat tcttaaaata aagtggtgat
     4921 cctaactgac ctaagacagg gaatttttac taggattaaa tgtcaggaat tgtgaaaaag
     4981 tgagtttaaa tgtatttggc taaggtgtat gtaaacttcc gacttcaact gtatagggat
     5041 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5101 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6961 tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7021 aaattgtaag cgttaatatt ttgttaaaat t
//
