LOCUS       sequence-224224-        6990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral plasmid containing mCerulean3 for labeling of cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6990)
  AUTHORS   Kleeman B, Olsson A, Newkold T, Kofron M, DeLay M, Hildeman D, 
            Grimes HL
  TITLE     A guide to choosing fluorescent protein combinations for flow 
            cytometric analysis based on spectral overlap.
  JOURNAL   Cytometry A. 2018 Mar 13. doi: 10.1002/cyto.a.23360.
  PUBMED    29533508
REFERENCE   2  (bases 1 to 6990)
  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: "Cytometry 
            A. 2018 Mar 13. doi: 10.1002/cyto.a.23360."
COMMENT     Sequence Label: pMSCVpuro-mCerulean3
FEATURES             Location/Qualifiers
     source          1..6990
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..39
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(77..99)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     199..218
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     412..434
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     562..581
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             781..1297
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1361..1702
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1769..2185
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2115..2137
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2153..2169
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2207..2216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        2946..3445
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2981..3004)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3378..3397
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             3466..4065
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3466..3485)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3962..3982
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4165..4679
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4841..4857
                     /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(4865..4871,4872..4889,4890..4895))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4910..4931
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5048..5065)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5219..5807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5299..5318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5978..6769,6770..6838))
                     /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     6601..6620
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6839..6943)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
       61 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
      121 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
      181 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt
      241 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
      301 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
      361 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      421 gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
      481 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
      541 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
      601 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa
      661 agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc agctgaagcc
      721 tatagagtac gagccataga taaaataaaa gattttattt agtctccaga aaaagggggg
      781 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
      841 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      901 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      961 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
     1021 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     1081 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     1141 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
     1201 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
     1261 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
     1321 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
     1381 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
     1441 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
     1501 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
     1561 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
     1621 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
     1681 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
     1741 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1801 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1861 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1921 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1981 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     2041 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     2101 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     2161 agccctcact ccttctctag gcgccggaat tagatctctc gaggttgcca ccatggtttc
     2221 taaaggggaa gaattgttta cgggagtggt gcctattttg gtggagcttg acggcgacgt
     2281 gaatgggcac aagttttccg tttctggtga aggagaaggg gatgccacat atggaaaact
     2341 caccctcaaa ttcatctgca ctacagggaa gctccctgtt ccgtggccaa cccttgtcac
     2401 cactctgtcc tggggcgttc agtgttttgc tagatatccc gatcatatga agcagcacga
     2461 ttttttcaaa tctgctatgc ccgaaggata cgtccaagaa cgaactattt ttttcaaaga
     2521 cgacgggaat tataaaacaa gggccgaggt gaagttcgag ggagacacac tcgtaaaccg
     2581 catcgaactc aaaggaatcg acttcaagga agacgggaat attctgggcc acaaactgga
     2641 atataacgcc attcacggca acgtgtacat tacagcagat aagcagaaaa acggaatcaa
     2701 agctaatttc ggtctgaatt gcaatattga agacggaagc gtgcagctcg ctgaccacta
     2761 tcagcagaac acccctatcg gcgatggacc tgtgctgctg ccggacaatc actacctgag
     2821 cacccagagt aagctgtcaa aggacccaaa tgagaagaga gatcacatgg ttctgctgga
     2881 gtttgttacc gctgcaggaa ttaccctcgg aatggatgag ctctacaaat gaaacgaatt
     2941 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
     3001 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
     3061 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
     3121 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
     3181 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
     3241 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
     3301 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
     3361 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     3421 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
     3481 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3541 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3601 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3661 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3721 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3781 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3841 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3901 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3961 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     4021 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
     4081 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cgataaaata aaagatttta
     4141 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     4201 taagtaacgc cattttgcaa ggcatggaaa atacataact gagaatagag aagttcagat
     4261 caaggttagg aacagagaga cagcagaata tgggccaaac aggatatctg tggtaagcag
     4321 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtccc gccctcagca
     4381 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     4441 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     4501 agctcaataa aagagcccac aacccctcac tcggcgcgcc agtcctccga tagactgcgt
     4561 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     4621 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatg
     4681 ggtaacagtt tcttgaagtt ggagaacaac attctgaggg taggagtcga atattaagta
     4741 atcctgactc aattagccac tgttttgaat ccacatactc caatactcct gaaatagttc
     4801 attatggaca gcgcagaaag agctggggag aattgtgaaa ttgttatccg ctcacaattc
     4861 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4921 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4981 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5041 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5101 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5161 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5221 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5281 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5341 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5401 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5461 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5521 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5581 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5641 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5701 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5761 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5821 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5881 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5941 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6001 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6061 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6121 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6181 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6241 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6301 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6361 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6421 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6481 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6541 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6601 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6661 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6721 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6781 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6841 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6901 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6961 tgccacctga cgtctaagaa accattatta
//
