LOCUS       sequence-341720-        8566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Firefly luciferase enhancer reporter plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8566)
  AUTHORS   Long HK, Osterwalder M, Welsh IC, Hansen K, Davies JOJ, Liu YE, 
            Koska M, Adams AT, Aho R, Arora N, Ikeda K, Williams RM, 
            Sauka-Spengler T, Porteus MH, Mohun T, Dickel DE, Swigut T, Hughes 
            JR, Higgs DR, Visel A, Selleri L, Wysocka J
  TITLE     Loss of Extreme Long-Range Enhancers in Human Neural Crest Drives a 
            Craniofacial Disorder.
  JOURNAL   Cell Stem Cell. 2020 Nov 5;27(5):765-783.e14. doi: 
            10.1016/j.stem.2020.09.001. Epub 2020 Sep 28.
  PUBMED    32991838
REFERENCE   2  (bases 1 to 8566)
  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"; doi: 
            "10.1016/j.stem.2020.09.001"; journalName: "Cell Stem Cell"; date: 
            "2020-11-5- 5"; volume: "27"; issue: "5"; pages: "765-783.e14"
COMMENT     Sequence Label: pGL3-noSV40-mouseEC1.45
FEATURES             Location/Qualifiers
     source          1..8566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3598..3794
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3645..3780
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3824..3833
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3830..5482
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3896..3914)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5523..5644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5534..5553)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5588..5607
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5898..5915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6069..6657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6149..6168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6828..7619,7620..7688))
                     /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     7451..7470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7689..7793)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7820..8275
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7907..7926)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8117..8138
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    8406..8454
                     /note="synthetic polyadenylation signal"
     misc_feature    8468..8559
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     8508..8527
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacctcca taacatcccc caaatggcac caacaaccaa gtgttctaat acccgagtct
       61 gtgtgggaca tttctcattc aaaccaccac agtaagtaaa ttaaatcatg cgaatgactt
      121 ttgtaaacta cgttgtgttg ggtaacgtaa ctgcaacata gcctagtttc tatttcacat
      181 caccaacatg attccatctg ataacgttca catagcatga tagcttctct actcatttca
      241 aggaagttca aagcatgttg aatttaactc tagcatgtaa atagtgcact cttctgtaat
      301 cctttagaat aatactacag tgccattgtt atttaatgtt aattacatca tttccttctt
      361 tctgatatca actctactta atcctgtctt acgccgctag ccagtgcctg gaggtgactg
      421 gcattgtgtt ttgcccagaa gttatagatg ctacttatga gagggtgagt gcttgtccaa
      481 atgaagtttg tcactcagag ctggaaattg aagcataaag atcaatacca aaacctccca
      541 gccactgctc cagaatacag tggagtttct tctgctccct gtttttattt atggtgtgta
      601 ctcatggaga aacaagccag ttccccccca ggacccagtg aagagcattt actcagtatt
      661 attggagcaa tggcatcgtt tttcagatat atccaagttc tgttccttga tctgtgacca
      721 ttatatttac ccaaaatatc cttctcatgg aaaacagttc tctggactct gcttgagact
      781 cataggctca gtcatctgct tcactttctc agaagagaag tcacatagtt acactaattc
      841 ttgctgcttc ttggcacccc caaattctct tagctaacga gagcttgtct tgggaccagg
      901 ctcttgggag ctttggagac catttcatcc acagagcttt ggaattccct cctcacccga
      961 catgtgcagc ttatacatca aacttgtggt ttgagacatt ttttttttca gatgcctgaa
     1021 gcatcaacag cactcttact ctaccagcat taacttatta ccatttcagt ggcatttgaa
     1081 gaagaaaagg agataaatac atacttgaaa gtgctgctac caatgttttt tttctacaca
     1141 tttgattgta tttggtgcaa ttatcggcat gtatctccac ttgccaagta taagagaggg
     1201 ttaggggtgt gtgtgtgtgt gtgtgtgtgt ccatccatct gtctgtgttt gtgtgtgatc
     1261 aaagatcttt gggggccatg ccctcattat ctttcagccc agcatctttg attaagaaca
     1321 ggtgcagcca agctctggcc ttgcctccca tttctgccgg catcaaataa aaagcagaag
     1381 tatgactgtt tgttggatct ttcccttact tttaattctc tggtgctgaa gaacatgcca
     1441 agtcatgggc aagaatctaa aattgtggct gaccaagggt cccacatccc ggcatgatcc
     1501 ccacagctct gcagttgtaa gataggaggt tttgatgtgg ttctgttgca cttctatctc
     1561 ccatgtaatt tcaatttaat cagcttccgt acatgctata aattatggct tccccgatct
     1621 atccttttta tgtgtatgcg gtgatgtaat tttctttttg agatttctat acttggatga
     1681 cagcatcctc ttatttctta accagataca aatgctcctg agtcggttct ttgtgtactc
     1741 cgaggactct gagattgatt gtgagttaaa tcatggaccc cgatgcccac ttcacacagg
     1801 tcagatcttt cacataatta ataacatgct atctactcta taattgggct ccacagaagc
     1861 atggtcactg tgatactgtt tgaccctcaa tcacagccta gcttatagat agctctattt
     1921 tccccatgtg caatgctttt ggtggattct gggatattat attgttggag tttcggtgag
     1981 cccagctatc tgaggacact ttactttgac acatgccact ttttagagga gagctggaaa
     2041 tgttcgttaa atgtgagcca aatggttagg gcagaaaagg caaaaccaaa gctaaatctt
     2101 tttttttttt ctttttcttt tttaaattta acataaatag agttgattct agggagataa
     2161 atgtattttt aatgatggga gaattctggg ttcgtacata aatcaaggca tcttgagttt
     2221 agagaaacag aaaatgagta atatcatatt tagctgtctt caccagagag tcctaattca
     2281 aaagtatgtc aaaagtggcc aggagtagct ctgagtgaaa gaatcatgga agtgcaacag
     2341 acagaaaccg gagaatggga aatgagagtc gaagaggttc cagaataacg tcgacgggag
     2401 ccaatgcaca gatctgtgtg ccaggaaaga tcggccccat ggagagagaa ctggatctga
     2461 ataaaggagc tatcagtacc aggacttaca tagacagagc attatgaagg aaggatacac
     2521 aacccttcca tgctccctgg gactgcttgg ccttaataag ctacaaagaa aattacaaag
     2581 atgaccagcc aaaggaaaaa ctcctttgca gactgtaggg gttattaatg aagatttaat
     2641 taaagattag tacttgaatt ggctcatgga caggtagaag aatatagatt tttttttaaa
     2701 ctggcttgaa ctcagatgaa atgtgttcat cttaattttg tatgacactc actacaaaat
     2761 ctccaaacta ctatcggtag aagttaggag aaattcccat aaataacatc ttctcgggag
     2821 atcagtagag aattttcatt ggaaagtgag gaacaaacca acaaacagac atgaactttc
     2881 ctcgattgta ttgcatttta aaagaaagga ttccaggaaa ccttcaggag acaccgcatg
     2941 ccttctgggg ccaggcagct gggagagaga gagggaagag attaacgtag aacaggtcgg
     3001 ggaccatccc aagcctatcc ttcaggccct cgattgaagg aattaagttt tattttatta
     3061 acttcgttct tgccctccca gagttgaggg tgctcactcc agggctcccg tagaaccgcc
     3121 ccagggaacc tcagtgcaaa aggtttgcac tcacagctgg ctgtgattac taggactggg
     3181 agagatcacg gcttgattgg ctttcaactg ctcttggttt gcttcagttc ctgcaggaca
     3241 attaatccac aggttcgagt aaatgttgga tcttaatgga taccagacac atcaggctca
     3301 accagaccac caaggctttg cggccccttt cctcacattt aattgtacaa atgaacttat
     3361 tcctcctcac ccaatggcca gaataacact gcttcgcggg ttttcttcat gacgttcatt
     3421 ctgtccttac tttgtcctta cttctgttcc atctgagtgt cgtcctcttc cgggtagtct
     3481 gccggcctcc ttgatcgtgt cttgagaggt cggaagttaa tcgtttgcac tcagggggat
     3541 gtgttggatg aaggacaaca catcctgcac tccttgtgct tctcgagatc tgcgatctgc
     3601 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3661 cgcccagttc cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg
     3721 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3781 taggcttttg caaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc
     3841 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
     3901 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
     3961 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
     4021 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     4081 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     4141 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     4201 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     4261 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     4321 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     4381 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     4441 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     4501 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     4561 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     4621 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     4681 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     4741 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     4801 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     4861 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     4921 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     4981 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     5041 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     5101 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     5161 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     5221 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     5281 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     5341 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     5401 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     5461 gggcggaaag atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat
     5521 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     5581 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     5641 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     5701 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc tggatccgtc
     5761 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     5821 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     5881 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5941 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6001 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6061 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6121 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6181 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6241 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6301 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6361 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6421 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6481 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6541 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6601 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6661 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6721 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6781 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6841 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6901 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6961 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7021 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7081 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7141 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7201 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7261 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7321 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7381 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7441 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7501 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7561 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7621 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7681 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7741 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7801 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     7861 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     7921 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     7981 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     8041 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     8101 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     8161 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     8221 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttgccat
     8281 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     8341 cgccagccca agctaccatg ataagtaagt aatattaagg tacgggaggt acttggagcg
     8401 gccgcaataa aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga
     8461 tagtactaac atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg
     8521 ctgtccccag tgcaagtgca ggtgccagaa catttctcta tcgata
//
