LOCUS       Exported                6838 bp DNA     circular SYN 17-MAR-2022
DEFINITION  corresponds to sh1 in the paper and sh2 in most data notes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6838)
  AUTHORS   Anna M. H. Hertle
  TITLE     Direct Submission
  JOURNAL   Exported Mar 17, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..190
                     /label=ITR
                     /note="color: #ffcc99"
     promoter        298..538
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     431..455
                     /label=U6_for
                     /note="color: #a020f0; direction: RIGHT"
     misc_RNA        550..570
                     /label=shNCLX-1 target sequence
                     /note="corresponds to nt 2389-2409 of Slc8b1 (NM_133221)"
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        629..1867
                     /label=CKIIa promoter
                     /note="color: #ffffff"
     primer_bind     complement(650..668)
                     /label=U6_rev
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     1722..1743
                     /label=CKIIa_for
                     /note="color: #a020f0; direction: RIGHT"
     CDS             1892..2602
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    2650..3238
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(2736..2754)
                     /label=WPRE_rev
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(3121..3132)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 3120"
                     /translation="IEGR"
     polyA_signal    3271..3495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3553..3734
                     /label=ITR
                     /note="color: #ffcc99"
     promoter        complement(3716..4045)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      3730..3865
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     rep_origin      complement(4291..4879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5050..5910)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5050 .. 5841 / #ccffcc
                        2: 5842 .. 5910 / #ccffcc / signal sequence
                     Cleavage site after base 5841"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5911..6015)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        6803..6821
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
ORIGIN
        1 tagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
       61 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      121 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      181 tctaggtacc ctcgaggagg agaagcatga attccccagt ggaaagacgc gcaggcaaaa
      241 cgcaccacgt gacggagcgt gaccgcgcgc cgagcgcgcg ccaaggtcgg gcaggaagag
      301 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
      361 attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
      421 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
      481 cttaccgtaa cttgaaagta tttcgatttc ttgggtttat atatcttgtg gaaaggacgc
      541 gggatcccca tgttggactg tggatctaaa ttcaagagat ttagatccac agtccaacat
      601 tttttggaaa agcttggcac tggccgtact agaaaatgag atgggggtgc agagagcttc
      661 ctcagtgacc tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta
      721 ctaatcttaa attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc
      781 attagggcag atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac
      841 cacccgccca gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca
      901 gtcctacctt gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc
      961 ttcatggtcc ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc
     1021 ctgctgctca gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt
     1081 ctgggtctgt ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta
     1141 aaaagcaggg agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca
     1201 attcccagag gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat
     1261 gaattagctt cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc
     1321 tgtctcttca gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca
     1381 tcccagaatg tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat
     1441 cccatcccgt ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg
     1501 atcacttgtg gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct
     1561 gggggaacag ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag
     1621 gccagggcaa agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt
     1681 accggggcaa cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac
     1741 gaaggctcgc gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag
     1801 tcaagccggt tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg
     1861 gggcagcggg ggatcaggtg gcagcggtgg catggtgagc aagggcgagg aggataacat
     1921 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
     1981 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     2041 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     2101 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     2161 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     2221 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     2281 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     2341 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     2401 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     2461 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     2521 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     2581 catggacgag ctgtacaagt agggtggcag cggtggcact cgagtaagct atcaagctag
     2641 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2701 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2761 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2821 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2881 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2941 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3001 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3061 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3121 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3181 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3241 cgataccgtc gactcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3301 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3361 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3421 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3481 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgact agagcatggc
     3541 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     3601 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     3661 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag agctttttgc
     3721 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     3781 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     3841 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     3901 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     3961 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     4021 actttccaca ccctaactga cacacattcc acagctgcat taatgaatcg gccaacgcgc
     4081 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5101 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5161 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6061 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6121 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6181 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6241 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattcgac
     6301 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     6361 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     6421 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     6481 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     6541 tgatgccggc cacgatgcgt ccggcgtaga ggatctggct agcgatgacc ctgctgattg
     6601 gttcgctgac catttccggg tgcgggacgg cgttaccaga aactcagaag gttcgtccaa
     6661 ccaaaccgac tctgacggca gtttacgaga gagatgatag ggtctgcttc agtaagccag
     6721 atgctacaca attaggcttg tacatattgt cgttagaacg cggctacaat taatacataa
     6781 ccttatgtat catacacata cgatttaggt gacactatag aatacacgga attaattc
//
