LOCUS       sequence-375724-        9060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Bcl11b .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9060)
  TITLE     Rothenberg Lab Plasmids 2022
REFERENCE   2  (bases 1 to 9060)
  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"
COMMENT     Sequence Label: Bcl11b MSCV IRES mCherry
FEATURES             Location/Qualifiers
     source          1..9060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1793..1811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1849..1871)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1971..1990
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2184..2206
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2334..2353
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2553..3069
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3133..3474
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3541..3957
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3887..3909
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3925..3941
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    6652..7219
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6669..7221
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6813..6830)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7040..7059
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      7242..7251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7248..7958
                     /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(7393..7411)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7661..7680
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             8007..8521
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    8683..8699
                     /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(8707..8713,8714..8731,8732..8737))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8752..8773
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8890..8907)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     1861 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     1921 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     1981 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     2041 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     2101 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     2161 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
     2221 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
     2281 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
     2341 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
     2401 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
     2461 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
     2521 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     2581 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     2641 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     2701 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     2761 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     2821 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     2881 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     2941 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     3001 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     3061 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
     3121 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
     3181 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
     3241 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     3301 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     3361 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     3421 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     3481 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     3541 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     3601 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     3661 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     3721 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     3781 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     3841 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     3901 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
     3961 attagatcta tgtcccgccg caaacagggc aacccgcagc acttgtccca gagggaactc
     4021 atcacgccag aggctgacca tgtggaggct accatcctcg aggaagacga gggtctggag
     4081 atagaggagc ctagcagcct ggggctgatg gtgggaggcc ccgaccctga tctactcacc
     4141 tgtggccagt gtcagatgaa cttcccgctg ggggacatcc tggtttttat agagcacaag
     4201 aagaaacagt gtggaggcct gggcccctgc tacgacaagg tcctggacaa gagcagtcca
     4261 cctccctcct ctcgctctga gctcaggaga gtatctgagc cagtggagat cgggatccag
     4321 gtcacccctg atgaagatga ccacctactg tcacccacga aaggcatctg tcccaagcag
     4381 gagaacattg cagggccgtg caggcctgcc cagctgccat cgatggcccc catagctgcc
     4441 tcctcttccc accctcccac ctccgtgatt acttcacctc tgcgtgccct gggcgttctc
     4501 ccaccctgtt tcccgctgcc ttgccgtggt gcacgcccca tctcgggcga cgggactcag
     4561 ggtgagggtc agatggaggc tccctttgga tgccagtgtg agttgtcagg taaagatgag
     4621 ccttccagct acatttgcac aacatgcaag cagcccttca acagcgcctg gttcctgctg
     4681 cagcacgcac agaacacaca tggcttccga atctacctgg agcctgggcc ggccagcacc
     4741 tcgctcacgc ccaggctcac catcccgcca ccgctcgggc cggagaccgt ggcgcagtcc
     4801 ccactcatga atttcctggg ggacagcaat cctttcaacc tgctgcgcat gacgggcccc
     4861 atcctgcggg accaccctgg cttcggtgag ggccgcttgc caggtacgcc accgctcttc
     4921 agcccaccgc cacgccatca cttggaccca caccgcctca gtgcagagga gatggggctc
     4981 gtggcccagc accccagtgc cttcgaccga gtcatgcgcc tgaaccccat ggccatagac
     5041 tctcctgcca tggacttctc ccggcggctg cgagaactgg ccggcaacag ctccacgccg
     5101 ccgcccgtgt ccccaggccg tggcaaccct atgcaccggc tgctgaaccc tttccagccc
     5161 agtcccaagt ccccgttcct cagcacgcca ccgctgccac ccatgcctgc gggcacaccg
     5221 ccaccgcagc cgcctgccaa gagcaagtcc tgtgagttct gcggcaagac cttcaagttc
     5281 cagagcaatc tcatcgtgca ccggcgcagc cacacgggcg agaagcccta caagtgccag
     5341 ctgtgcgacc atgcgtgctc gcaggcgagc aagctcaagc gccacatgaa gacgcacatg
     5401 cacaaggcgg gctctctggc tggccgctca gacgacgggc tctcagctgc cagctcccct
     5461 gagccgggca ccagcgagct gccaggtgac ctgaaagcgg ccgatggcga cttccgccac
     5521 catgagagcg acccatctct gggccccgag cctgaggacg acgaggacga ggaggaggaa
     5581 gaagaggagc tgctgctgga gaacgagagc cggcctgagt cgagcttcag catggactcg
     5641 gagctgggcc gtggccgcga gaacggaggt ggcgtgccac cgggggtggc gggcgcaggg
     5701 gctgcagctg cggctctggc ggatgagaag gctctggccc tgggcaaggt gatggaggac
     5761 gcagggctgg gcgcactgcc gcagtatggg gagaagcggg gcgccttcct gaagcgtgca
     5821 ggcgacacgg gtgatgccgg agctgttggc tgtggggacg cgggtgcacc gggtgcagtg
     5881 aacgggcgcg gcggggcctt cgcgccaggc gcagagccct ttccagctct cttcccacgc
     5941 aagccagcac cgctgcccag ccctgggctc ggtggtcccg cgctgcacgc ggccaagcgc
     6001 atcaaggtgg agaaagacct ggagctgcca cctgccgccc tcatcccatc tgagaacgtg
     6061 tactcgcagt ggctcgtggg ctacgcagca tcgcgccact tcatgaagga cccattcctg
     6121 ggcttcacgg atgcgcgcca gtcgcctttc gccacatcgt cggaacattc ctctgagaac
     6181 ggcagcctgc gcttctcaac gccacccggg gacctgctgg acggcgggct gtccgggcgc
     6241 agtggcacgg cgagcggggg cagcacacct cacctgggtg gtccgggtcc tgggaggccg
     6301 agctccaagg agggccgccg cagcgacaca tgtgagtact gcggcaaggt cttcaagaac
     6361 tgtagcaacc tgacggtgca ccggaggagc cacaccggcg agcggcctta caagtgcgag
     6421 ctgtgcaact acgcgtgcgc gcagagcagc aagctcacgc gccacatgaa gacgcacggg
     6481 cagatcggca aggaggtgta ccgctgcgac atctgccaga tgcccttcag cgtctacagc
     6541 accctggaga aacacatgaa aaagtggcac ggtgaacact tgctgactaa tgatgtcaaa
     6601 atcgagcagg ctgagaggag ctaagaattc tgctatacga agttatccct ctccctcccc
     6661 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     6721 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     6781 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     6841 gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc
     6901 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     6961 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     7021 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     7081 ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat gctttacatg
     7141 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     7201 ttgaagaaca cgatgataat atggccacaa ccatgactag tgccaccatg gtgagcaagg
     7261 gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg
     7321 gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg
     7381 gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca
     7441 tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc
     7501 ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg
     7561 aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct
     7621 acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga
     7681 ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg
     7741 agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca
     7801 cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg
     7861 acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc
     7921 gccactccac cggcggcatg gacgagctgt acaagtaagc atcgataaaa taaaagattt
     7981 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     8041 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     8101 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     8161 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     8221 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     8281 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     8341 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     8401 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     8461 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     8521 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
     8581 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
     8641 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     8701 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     8761 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     8821 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     8881 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     8941 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     9001 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
//
