LOCUS       Exported                6834 bp DNA     circular SYN 17-MAR-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6834)
  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..6834
                     /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..568
                     /label=shCTRL
                     /note="no known targets in the mouse genome"
                     /note="color: #993300; direction: RIGHT"
     promoter        625..1863
                     /label=CKIIa promoter
                     /note="color: #ffffff"
     primer_bind     complement(646..664)
                     /label=U6_rev
                     /note="color: #a020f0; direction: LEFT"
     primer_bind     1718..1739
                     /label=CKIIa_for
                     /note="color: #a020f0; direction: RIGHT"
     CDS             1888..2598
                     /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    2646..3234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(2732..2750)
                     /label=WPRE_rev
                     /note="color: #a020f0; direction: LEFT"
     CDS             complement(3117..3128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 3116"
                     /translation="IEGR"
     polyA_signal    3267..3491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   3549..3730
                     /label=ITR
                     /note="color: #ffcc99"
     promoter        complement(3712..4041)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      3726..3861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     rep_origin      complement(4287..4875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5046..5906)
                     /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: 5046 .. 5837 / #ccffcc
                        2: 5838 .. 5906 / #ccffcc / signal sequence
                     Cleavage site after base 5837"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5907..6011)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     promoter        6799..6817
                     /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 gggatctccg tgccaagacg ggtagtcatt caagagatga ctacccgtct tggcactttt
      601 tggaagagct tggcactggc cgtactagaa aatgagatgg gggtgcagag agcttcctca
      661 gtgacctgcc cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa
      721 tcttaaattc catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta
      781 gggcagatga ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc
      841 cgcccagccc tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc
      901 taccttgacg ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca
      961 tggtccctct gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc
     1021 tgctcagtgg tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg
     1081 gtctgtttgg tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa
     1141 gcagggagag ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc
     1201 ccagaggaag caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat
     1261 tagcttcccc tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc
     1321 tcttcagagg agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc
     1381 agaatgtggc acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca
     1441 tcccgtctgt taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca
     1501 cttgtggact aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg
     1561 gaacagggtc cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca
     1621 gggcaaagag gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg
     1681 gggcaacggg aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag
     1741 gctcgcgagg ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa
     1801 gccggttctc cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc
     1861 agcgggggat caggtggcag cggtggcatg gtgagcaagg gcgaggagga taacatggcc
     1921 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1981 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     2041 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     2101 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     2161 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     2221 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     2281 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     2341 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     2401 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
     2461 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
     2521 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
     2581 gacgagctgt acaagtaggg tggcagcggt ggcactcgag taagctatca agctagctta
     2641 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3241 accgtcgact cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3301 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3361 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3421 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3481 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag catggctacg
     3541 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     3601 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3661 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagagct ttttgcaaaa
     3721 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3781 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3841 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     3901 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     3961 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4021 tccacaccct aactgacaca cattccacag ctgcattaat gaatcggcca acgcgcgggg
     4081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5041 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5101 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5161 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5221 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5281 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5341 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5401 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5461 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5521 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5581 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5641 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5701 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5761 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5821 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5881 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5941 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6001 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     6061 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     6121 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     6181 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     6241 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca ttcgacgctc
     6301 tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac
     6361 cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg
     6421 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     6481 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6541 gccggccacg atgcgtccgg cgtagaggat ctggctagcg atgaccctgc tgattggttc
     6601 gctgaccatt tccgggtgcg ggacggcgtt accagaaact cagaaggttc gtccaaccaa
     6661 accgactctg acggcagttt acgagagaga tgatagggtc tgcttcagta agccagatgc
     6721 tacacaatta ggcttgtaca tattgtcgtt agaacgcggc tacaattaat acataacctt
     6781 atgtatcata cacatacgat ttaggtgaca ctatagaata cacggaatta attc
//
