LOCUS       sequence-139261-        5852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Crx shRNA (mir30 backbone) was inserted into the 3'UTR of mCherry..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  AUTHORS   Wang S, Sengel C, Emerson MM, Cepko CL
  TITLE     A gene regulatory network controls the binary fate decision of rod 
            and bipolar cells in the vertebrate retina.
  JOURNAL   Dev Cell. 2014 Sep 8;30(5):513-27. doi: 
            10.1016/j.devcel.2014.07.018. Epub 2014 Aug 21.
  PUBMED    25155555
REFERENCE   2  (bases 1 to 5852)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.devcel.2014.07.018"; journalName: "Dev Cell"; date: 
            "2014-09-8- 8"; volume: "30"; issue: "5"; pages: "513-27"
COMMENT     Sequence Label: CAG-mCherry-CrxshRNA
FEATURES             Location/Qualifiers
     source          1..5852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1725..2435
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1870..1888)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2138..2157
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           2468..2589
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2617..2631
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2659..2787
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(2873..2892)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2938..2993
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2992..3011)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3354..3370)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3354..3370)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3367..3389)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3378..3394
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3402..3408,3409..3426,3427..3432))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3448..3469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3528..3723
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3574..3709
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3729..3863
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3766..3785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3820..3839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3931..3948)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4102..4690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4182..4201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4861..5652,5653..5721))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVXAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5484..5503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5722..5826)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgatggtg agcaagggcg
     1741 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1801 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1861 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1921 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1981 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     2041 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     2101 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2161 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     2221 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     2281 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     2341 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2401 actccaccgg cggcatggac gagctgtaca agtaagcggc cgcgactagg gataacaggg
     2461 taattgtttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat cttggaaaca
     2521 cttgctggga ttacttcttc aggttaaccc aacagaaggc tcgagaaggt atattgctgt
     2581 tgacagtgag cgaaccaggc tgtcccatac tcaatagtga agccacagat gtattgagta
     2641 tgggacagcc tggtgtgcct actgcctcgg agaattcaag gggctacttt aggagcaatt
     2701 atcttgttta ctaaaactga ataccttgct atctctttga tacattttta caaagctgaa
     2761 ttaaaatggt ataaattaaa tcactttgcg gccgcactcc tcaggtgcag gctgcctatc
     2821 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     2881 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2941 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     3001 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     3061 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     3121 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3181 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     3241 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     3301 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     3361 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3421 ataaagtgta aagcctgggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3481 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     3541 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3601 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3661 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3721 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3781 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3841 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     3901 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3961 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4021 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4081 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4141 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4201 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4261 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     4321 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4441 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4501 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     4561 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4621 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4681 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4741 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4801 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4861 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4921 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4981 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5041 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5101 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5161 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5221 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5281 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5341 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5401 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5461 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5521 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5581 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5641 cgtttctggg tgagcasaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5701 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5761 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5821 tccgcgcaca tttccccgaa aagtgccacc tg
//
