LOCUS       sequence-337469-        8106 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the expression of Luciferase and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8106)
  TITLE     Saradhi Mallampati (Pre-published)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8106)
  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: 
            "Unpublished"
COMMENT     Sequence Label: MIG-Luciferase-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8106
                     /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
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1436..1445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1442..3094
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    3099..3666
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3116..3668
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3260..3277)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3487..3506
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3680..3682,3683..3685,3686..4399)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3725..3746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3986..4005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4333..4354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4501..5015
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5177..5193
                     /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(5201..5207,5208..5225,5226..5231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5246..5267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5384..5401)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5555..6143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5635..5654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6314..7105,7106..7174))
                     /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     6937..6956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7175..7279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7347..7365
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7403..7425)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7525..7544
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7738..7760
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7888..7907
                     /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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg
     3781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     3841 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     3901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     3961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4081 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4141 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     4201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4261 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4381 ggacgagctg tacaagtaat gaattaatta agaattatca agcttatcga taccgtcgac
     4441 ctgcagccaa gcttatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     4501 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     4561 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
     4621 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     4681 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
     4741 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     4801 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     4861 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat
     4921 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     4981 gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag
     5041 aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt
     5101 ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct
     5161 ggggagaatt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5221 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5281 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5341 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5401 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5461 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6181 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6241 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6301 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6361 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6421 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6481 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6541 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6601 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6661 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6721 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6781 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6841 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6901 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6961 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7021 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7081 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7141 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7201 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7261 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7321 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7381 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7441 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7501 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7561 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     7621 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7681 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     7741 agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa
     7801 cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc
     7861 gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc
     7921 acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt
     7981 tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct
     8041 gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa
     8101 gggggg
//
