LOCUS       Exported                6625 bp ds-DNA     circular SYN 23-SEP-2022
DEFINITION  SB-transposon with constitutive bi-directional promoter, one side: 
            SfiI cloning site for GOI (contains filler DNA), other side: RFP and
            puromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    pSBbi-RP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6625)
  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 6625)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6625
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    410..634
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     623..640
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      880..889
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(915..2096)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(947..967)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2729..3430
                     /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(2862..2880)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3130..3149
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3324..3347
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             3440..3496
                     /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             3497..4096
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3497..3516)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3993..4013
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(4196..4215)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4261..4316
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4315..4334)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4708..4725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4879..5467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4959..4978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5638..6498)
                     /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     6261..6280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6499..6603)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
       61 gtaagcgtta atattttgtt aaaattggat ccctatacag ttgaagtcgg aagtttacat
      121 acacttaagt tggagtcatt aaaactcgtt tttcaactac tccacaaatt tcttgttaac
      181 aaacaatagt tttggcaagt cagttaggac atctactttg tgcatgacac aagtcatttt
      241 tccaacaatt gtttacagac agattatttc acttataatt cactgtatca caattccagt
      301 gggtcagaag tttacataca ctaagttcga ctcctctgca gaatgcggcg atgtttcggt
      361 aaggggtccg ctatctagta ggcccagctg gttctttccg cctcagaagc catagagccc
      421 accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct gtcctgcccc
      481 accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat ttcctcattt
      541 tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac gggggagggg
      601 caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg agctctagag
      661 aattgatccc caagcttggc ctgacaggcc ttcgaattag tgagaaggaa agacgcaacc
      721 aggaagccca ccaagagaaa gaggcatttc ctgaaaagat tccccttttt ggagagccct
      781 acaaggtatt tactgaacac tagacattga agtcctgatt tatcacaatg ttgaacccta
      841 tgatgaaaca attcagtata attgagttcg aaactcgagc catggtggcc tcagaggcca
      901 gcttggggta gttttcacga caccttaaat ggaagaaaaa aactttgaac cactgtctga
      961 ggcttgagaa tgaaccaaga tccaaactca aaaagggcaa attccaagga gaattacatc
     1021 aagtgccaag ctggcctaac ttcagtctcc acccactcag tatggggaaa ctccatcgca
     1081 taaaacccct ccccccaacc taaagacgac gtactccaaa agctcgagaa ctaatcgagg
     1141 tgcctggacg gcgcccggta ctccgtggag tcacatgaag cgacggctga ggacggaaag
     1201 gcccttttcc tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg cgtcctccat
     1261 tttgagctcc ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc aactggtgcc
     1321 gaccgggcca gccttgccgc ccagggcggg gcgatacacg gcggcgcgag gccaggcacc
     1381 agagcaggcc ggccagcttg agactacccc cgtccgattc tcggtggccg cgctcgcagg
     1441 ccccgcctcg ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc cgcggcccca
     1501 aaaaccgaaa taccagtgtg cagatcttgg cccgcattta caagactatc ttgccagaaa
     1561 aaaagcgtcg cagcaggtca tcaaaaattt taaatggcta gagacttatc gaaagcagcg
     1621 agacaggcgc gaaggtgcca ccagattcgc acgcggcggc cccagcgccc aagccaggcc
     1681 tcaactcaag cacgaggcga aggggctcct taagcgcaag gcctcgaact ctcccaccca
     1741 cttccaaccc gaagctcggg atcaagaatc acgtactgca gccaggggcg tggaagtaat
     1801 tcaaggcacg caagggccat aacccgtaaa gaggccaggc ccgcgggaac cacacacggc
     1861 acttacctgt gttctggcgg caaacccgtt gcgaaaaaga acgttcacgg cgactactgc
     1921 acttatatac ggttctcccc caccctcggg aaaaaggcgg agccagtaca cgacatcact
     1981 ttcccagttt accccgcgcc accttctcta ggcaccggtt caattgccga cccctccccc
     2041 caacttctcg gggactgtgg gcgatgtgcg ctctgcccac tgacgggcac cggagcctct
     2101 aggcgagacc ctgtctcaca aaataaagta agcccggact gagtgcggaa aggcgggcct
     2161 ggcgggtctg gtctccccat gcgggccacc agaggccctg cagccttcag tcgcttgaag
     2221 gggtaatggc gcttccactc acaaacatgg cggacagagc gtgtgaacga gatgaacagc
     2281 ccctcaaaaa tatggccgcc gaggctggac ggccgtgccc cagcagcacc gcctccgcgc
     2341 cccacgtgat ctctcgccgg gcacagcgct gaccgcggag gtccaaccgg aagaatgtcc
     2401 ggattggaca ttcggaagag ggcccgcctt ccctggggaa tctctgcgca cgcgcagaac
     2461 gcttcgacca atgaaaacac aggaagccgt ccgcgcaacc gcgttgcgtc acttctgccg
     2521 cccctgtttc aaggtatata gccgtagacg gaacttcgcc tttctctcgg ccttagcgcc
     2581 atttttttgg gtgagtgttt tttggttcct gcgttgggat tccgtgtaca atccatagac
     2641 atctgacctc ggcacttagc atcatcacag caaactaact gtagcctttc tctctttccc
     2701 tgtagaaacc tctgcacctg aggccaccat gttgagcaag ggcgaggagg tcatcaaaga
     2761 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccatgagt tcgagatcga
     2821 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2881 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
     2941 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
     3001 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
     3061 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
     3121 ccccgacggc cccgtaatgc agaagaagac aatgggctgg gaggcctcca ccgagcgcct
     3181 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
     3241 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
     3301 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
     3361 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctgtacg gcatggacga
     3421 gctgtacaag ggcagtggag ctactaactt cagcctgctg aagcaggctg gtgacgtcga
     3481 ggagaatcct ggccccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     3541 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     3601 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     3661 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     3721 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     3781 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     3841 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     3901 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     3961 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     4021 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     4081 caagcccggt gcctgattcg aaggcctgtc gtgaattcac tcctcaggtg caggctgcct
     4141 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     4201 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     4261 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     4321 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     4381 tggcaacata tgccatatgc tggctgccat gaactagcta ctcgggaccc cttaccgaaa
     4441 catcgccgca ttctgcagag gagtcgagtg tatgtaaact tctgacccac tgggaatgtg
     4501 atgaaagaaa taaaagctga aatgaatcat tctctctact attattctga tatttcacat
     4561 tcttaaaata aagtggtgat cctaactgac ctaagacagg gaatttttac taggattaaa
     4621 tgtcaggaat tgtgaaaaag tgagtttaaa tgtatttggc taaggtgtat gtaaacttcc
     4681 gacttcaact gtatagggat ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4741 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4801 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4861 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4921 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4981 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5041 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5101 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5161 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5221 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5281 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5341 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5401 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5461 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5521 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5581 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5641 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5701 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5761 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5821 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5881 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5941 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6001 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6061 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6121 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6181 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6241 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6301 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6361 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6421 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6481 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6541 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6601 gcgcacattt ccccgaaaag tgcca
//
