LOCUS       40924_1954        8086 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Encodes the expression of Luciferase and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  AUTHORS   Mallampati S, Sun B, Lu Y, Ma H, Gong Y, Wang D, Lee JS, Lin K, Sun 
            X
  TITLE     Integrated genetic approaches identify the molecular mechanisms of 
            Sox4 in early B-cell development: intricate roles for RAG1/2 and 
            CK1epsilon.
  JOURNAL   Blood. 2014 Jun 26;123(26):4064-76. doi: 
            10.1182/blood-2013-12-543801. Epub 2014 Apr 30.
  PUBMED    24786772
REFERENCE   2  (bases 1 to 8086)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8086)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1182/blood-2013-12-543801"; journalName: "Blood"; date: 
            "2014-06-26- 26"; volume: "123"; issue: "26"; pages: "4064-76"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             989..1405
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     CDS             1442..3091
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    3103..3679
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3701..4408
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     LTR             4481..4995
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     protein_bind    complement(5157..5173)
                     /label=lac operator
                     /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(5181..5211)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5226..5247)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(5364..5381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5535..6123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6297..7154)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7155..7259)
                     /label=AmpR promoter
     primer_bind     7327..7345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     primer_bind     complement(7383..7405)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7505..7524
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     7718..7740
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7868..7887
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI,
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctctc gaggttaacg aattcgccac
     1441 catggaagat gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg
     1501 gaccgccggc gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat
     1561 cgcctttacc gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag
     1621 cgttcggctg gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt
     1681 gtgcagcgag aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt
     1741 ggctgtggcc ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat
     1801 cagccagccc accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca
     1861 aaagaagcta ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg
     1921 cttccaaagc atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga
     1981 cttcgtgccc gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg
     2041 cagtaccgga ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag
     2101 tcatgcccgc gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt
     2161 ggtgccattt caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt
     2221 tcgggtcgtg ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta
     2281 taagattcaa tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct
     2341 catcgacaag tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag
     2401 caaggaggta ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta
     2461 cggcctgaca gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg
     2521 cgcagtaggc aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa
     2581 gacactgggt gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg
     2641 ctacgttaac aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag
     2701 cggcgacatc gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag
     2761 cctgatcaaa tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca
     2821 acaccccaac atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct
     2881 gcccgccgca gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga
     2941 ctatgtggcc agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga
     3001 cgaggtgcct aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat
     3061 taaggccaag aagggcggca agatcgccgt gtaagaattc cgcccccccc ccctaacgtt
     3121 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     3181 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     3241 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     3301 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     3361 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     3421 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     3481 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     3541 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     3601 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     3661 tgataatatg gccacaacca tggatccacc ggtcgccacc atggtgagca agggcgagga
     3721 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     3781 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3841 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     3901 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     3961 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     4021 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     4081 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     4141 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     4201 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     4261 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     4321 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     4381 caccggcggc atggacgagc tgtacaagta gcggcccgac ctgcagccaa gcttatcgat
     4441 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     4501 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     4561 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     4621 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     4681 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     4741 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     4801 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     4861 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     4921 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     4981 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     5041 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     5101 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     5161 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5221 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5281 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5341 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5401 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5461 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5521 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5581 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5641 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5701 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     5761 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5821 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5881 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5941 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6001 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6061 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6121 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6181 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6241 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6301 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6361 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6421 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6481 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6541 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6601 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6661 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6721 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6781 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6841 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6901 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6961 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7021 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7081 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7141 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7201 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7261 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     7321 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     7381 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     7441 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     7501 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     7561 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     7621 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     7681 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     7741 acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     7801 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     7861 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     7921 cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac aggatatcag
     7981 tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata gagtacgagc
     8041 catagataaa ataaaagatt ttatttagtc tccagaaaaa gggggg
//
