LOCUS       Exported File           6820 bp ds-DNA    circular SYN 29-SEP-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pSBbi_RN
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6820)
  AUTHORS   Eric Kowarz
  TITLE     .
  JOURNAL   .
FEATURES             Location/Qualifiers
     source          1..6820
                     /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..4205
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4223..4523
                     /note="rBG-pA"
                     /note="rabbit beta-globin polyadenylation signal"
                     /note="color: #cc0000; direction: RIGHT"
     misc_feature    4534..4575
                     /note="3'-proxITR"
                     /note="modified region proximal to 3'-ITR - contains CTCF 
                     binding sites"
                     /note="color: #cc0000"
     misc_feature    4576..4800
                     /note="3'-ITR"
                     /note="Sleeping Beauty inverted tandem repeat"
                     /note="color: #cc00cc"
     misc_feature    4801..4804
                     /note="tata"
                     /note="color: #cc0000"
     rep_origin      complement(4988..5576)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5747..6607)
                     /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: 5747 .. 6538 / #ccffcc
                        2: 6539 .. 6607 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6608..6712)
                     /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 atgattgaac
     3421 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3481 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3541 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3601 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3661 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3721 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3781 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3841 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3901 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3961 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4021 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4081 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4141 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4201 tctgattcga aggcctgtcg tgaattcact cctcaggtgc aggctgccta tcagaaggtg
     4261 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     4321 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     4381 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     4441 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     4501 gccatatgct ggctgccatg aactagctac tcgggacccc ttaccgaaac atcgccgcat
     4561 tctgcagagg agtcgagtgt atgtaaactt ctgacccact gggaatgtga tgaaagaaat
     4621 aaaagctgaa atgaatcatt ctctctacta ttattctgat atttcacatt cttaaaataa
     4681 agtggtgatc ctaactgacc taagacaggg aatttttact aggattaaat gtcaggaatt
     4741 gtgaaaaagt gagtttaaat gtatttggct aaggtgtatg taaacttccg acttcaactg
     4801 tatagggatc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4861 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4921 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4981 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5041 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5101 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5161 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5221 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5281 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5341 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5401 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5461 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5521 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5581 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5641 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5701 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5761 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5821 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5881 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5941 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6001 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6061 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6121 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6181 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6241 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6301 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6361 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6421 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6481 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6541 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6601 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6661 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6721 cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6781 cgcatcagga aattgtaagc gttaatattt tgttaaaatt
//