LOCUS       sequence-171275-        6807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Short hairpin RNAs were designed to target murine Cit (encoding 
            Citron Rho-interacting kinase) and were subcloned into the 
            Banshee-GFP vector for expression under control of the H1 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6807)
  AUTHORS   Swartz KL, Wood SN, Murthy T, Ramirez O, Qin G, Pillai MM, Rao S, 
            Minella AC
  TITLE     E2F-2 promotes nuclear condensation and enucleation of terminally 
            differentiated erythroblasts.
  JOURNAL   Mol Cell Biol. 2016 Oct 17. pii: MCB.00274-16.
  PUBMED    27795297
REFERENCE   2  (bases 1 to 6807)
  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"; journalName: "Mol Cell 
            Biol. 2016 Oct 17. pii: MCB.00274-16."
COMMENT     Sequence Label: Banshee-ShCit-B
FEATURES             Location/Qualifiers
     source          1..6807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(1176..1192)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1176..1192)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1229..1247)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1268..1284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1268..1284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1281..1303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1292..1308
                     /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(1316..1322,1323..1340,1341..1346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1361..1382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1499..1516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1750..1769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2429..3220,3221..3289))
                     /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     3052..3071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3290..3394)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3420..3875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3557..3578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3769..3788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4002..4024
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4016..4033
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4017..4033
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4040..4059
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4040..4058
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             4521..4696
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    4760..5101
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             5168..5584
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     5514..5536
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     5552..5568
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      5630..5639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5636..5638,5639..5641,5642..6355)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5636..5638,5639..5641,5642..6352)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5681..5702)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5942..5961)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6289..6310
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6466..6485
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc tgtgcgccgt gtgtctggat
      241 ttcaagagaa tccagacaca cggcgcactt tttaagcttg gatcaaggtc agaaacagag
      301 agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
      361 ggccaagaac agttggaaca gcagaatatg ggccaaacag gatatctgtg gtaagcagtt
      421 cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc cctcagcagt
      481 ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
      541 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
      601 ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg
      661 cccgggtacc cgtgttctca ataaaccctc ttgcagttgc atccgactcg tggtctcgct
      721 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcagttt
      781 ctcccaccta cacaggtctc actaacattc ctgatgtgcc gcagggactc cgtcagcccg
      841 gtttttgttt ataataaaat gcaagaacag tgttcccttc aagccagact acatcctgac
      901 tctcggcttt ataaaagaat gttgaagggc tctgtggact atctgccaca cgacttttaa
      961 gatttttatg cctcctggat gagggattta gtcaatctat cctcgtctat tttgctggct
     1021 tctccgtatt ttaaatttct agtttgcact cccttcctga gagcacggcg attgcagagt
     1081 agttaatact ctgagggcag gcttctgtga aaaggttgcc tgggctcagt gtgagatttt
     1141 gccataaaaa ggggtcctgc ccctgtgtac agacagatcc actagttcta gagcggccgc
     1201 caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
     1261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3421 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3481 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3541 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3601 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3661 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3721 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3781 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     3841 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     3901 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3961 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     4021 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggta ccgggccccc
     4081 ccgccgcggc ccttccgttt ctttgctttt gaaagacccc acccgtaggt ggcaagctag
     4141 cttaagtaac gccactttgc aaggcatgga aaaatacata actgagaata gaaaagttca
     4201 gatcaaggtc aggaacaaag aaacagctga ataccaaaca ggatatctgt ggtaagcggt
     4261 tcctgccccg gctcagggcc aagaacagat gagacagctg agtgatgggc caaacaggat
     4321 atctgtggta agcagttcct gccccggctc ggggccaaga acagatggtc cccagatgcg
     4381 gtccagccct cagcagtttc tagtgaatca tcagatgttt ccagggtgcc ccaaggacct
     4441 gaaaatgacc ctgtacctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     4501 gcgcttccgc tctccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     4561 ttccgataga ctgcgtcgcc cgggtacccg tattcccaat aaagcctctt gctgtttgca
     4621 tccgaatcgt ggactcgctg atccttggga gggtctcctc agattgattg actgcccacc
     4681 tcgggggtct ttcatttgga ggttccaccg agatttggag acccctgccc agggaccacc
     4741 gacccccccg ccgggaggta agctggccag cggtcgtttc gtgtctgtct ctgtctttgt
     4801 gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg tactagttag ctaactagct
     4861 ctgtatctgg cggacccgtg gtggaactga cgagttctga acacccggcc gcaaccctgg
     4921 gagacgtccc agggactttg ggggccgttt ttgtggcccg acctgaggaa gggagtcgat
     4981 gtggaatccg accccgtcag gatatgtggt tctggtagga gacgagaacc taaaacagtt
     5041 cccgcctccg tctgaatttt tgctttcggt ttggaaccga agccgcgcgt cttgtctgct
     5101 gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga
     5161 aaattagggc cagactgtta ccactccctt aagtttgacc ttaggtcact ggaaagatgt
     5221 cgagcggatc gctcacaacc agtcggtaga tgtcaagaag agacgttggg ttaccttctg
     5281 ctctgcagaa tggccaacct ttaacgtcgg atggccgcga gacggcacct ttaaccgaga
     5341 cctcatcacc caggttaaga tcaaggtctt ttcacctggc ccgcatggac acccagacca
     5401 ggtcccctac atcgtgacct gggaagcctt ggcttttgac ccccctccct gggtcaagcc
     5461 ctttgtacac cctaagcctc cgcctcctct tcctccatcc gccccgtctc tcccccttga
     5521 acctcctcgt tcgaccccgc ctcgatcctc cctttatcca gccctcactc cttctctagg
     5581 cgccggaatt ctgcagtcga cggtaccgcg ggcccgggat ccaccggtcg ccaccatggt
     5641 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     5701 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     5761 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     5821 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     5881 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     5941 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     6001 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     6061 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     6121 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     6181 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     6241 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     6301 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     6361 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     6421 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     6481 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     6541 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     6601 gactctgggg ttcggtagtc gacggatcct ctagacgcgt ctcgagtcga cggtatcgat
     6661 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     6721 atcacagctg aagcctatag agtacgagcc atagatagaa taaaagattt tatttagtct
     6781 ccagaaaaag gggggaatga aagatcc
//
