LOCUS       sequence-341793-        5121 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 5121)
  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 5121)
  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-humanCOL2A1enhancer
FEATURES             Location/Qualifiers
     source          1..5121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      65..200
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             250..1902
                     /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(316..334)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1943..2064
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1954..1973)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2008..2027
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2318..2335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2489..3077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2569..2588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3248..4039,4040..4108))
                     /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     3871..3890
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4109..4213)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4240..4695
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4327..4346)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4537..4558
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4826..4874
                     /note="synthetic polyadenylation signal"
     misc_feature    4888..4979
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4928..4947
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 gctcgagatc tgcgatctgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
       61 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatc gctgactaat
      121 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
      181 aggaggcttt tttggaggcc taggcttttg caaaaagctt ggcattccgg tactgttggt
      241 aaagccacca tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg
      301 gaagatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct
      361 ggaacaattg cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc
      421 gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga
      481 atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt
      541 atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt
      601 atgggcattt cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg
      661 aacgtgcaaa aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat
      721 taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat
      781 gaatacgatt ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac
      841 tcctctggat ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg
      901 agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt
      961 ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata
     1021 tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt
     1081 caggattaca agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa
     1141 agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct
     1201 cccctctcta aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg
     1261 caaggatatg ggctcactga gactacatca gctattctga ttacacccga gggggatgat
     1321 aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat
     1381 accgggaaaa cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt
     1441 atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg
     1501 ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc
     1561 ctgaagtctc tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc
     1621 ttgctccaac accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc
     1681 ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag
     1741 atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg
     1801 tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag
     1861 atcctcataa aggccaagaa gggcggaaag atcgccgtgt aattctagag tcggggcggc
     1921 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1981 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2041 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     2101 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
     2161 gataaggatc tggatccgtc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
     2221 ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct ttatcatgca
     2281 actcgtagga caggtgccgg cagcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2341 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2401 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2461 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2521 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2581 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2641 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2701 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2761 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2821 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2881 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2941 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3001 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3061 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3121 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3181 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3241 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3301 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3361 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3421 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3481 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3541 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3601 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3661 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3721 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3781 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3841 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3901 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3961 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4021 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4081 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4141 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4201 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     4261 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4321 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4381 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4441 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4501 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4561 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4621 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4681 taacgcttac aatttgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4741 tgcgggcctc ttcgctatta cgccagccca agctaccatg ataagtaagt aatattaagg
     4801 tacgggaggt acttggagcg gccgcaataa aatatcttta ttttcattac atctgtgtgt
     4861 tggttttttg tgtgaatcga tagtactaac atacgctctc catcaaaaca aaacgaaaca
     4921 aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca ggtgccagaa tatttctcta
     4981 tcgataggta cccttccaga tggggctgaa acgctgcccg tatttattta aactggttcc
     5041 tcgcggagac ctgtgaatcg ggctctgtgt gcgctcgaga aaagccccat tcatgagaga
     5101 cgaggtccag tgggttctct c
//
