LOCUS       sequence-341750-        6938 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 6938)
  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 6938)
  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-humanEC1.35_S1-2
FEATURES             Location/Qualifiers
     source          1..6938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1970..2166
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2017..2152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2196..2205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2202..3854
                     /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(2268..2286)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3895..4016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3906..3925)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3960..3979
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4270..4287)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4441..5029)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4521..4540)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5200..5991,5992..6060))
                     /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     5823..5842
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6061..6165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6192..6647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6279..6298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6489..6510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6778..6826
                     /note="synthetic polyadenylation signal"
     misc_feature    6840..6931
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6880..6899
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtacctggg atgagagaga aagggctgtt ttgtctctca aggaattgct aaaccagatg
       61 ttgaaaaggg aggagagatt tttaaatttt agttgaaaag cagtatccaa tgaattaaaa
      121 actgataccc aatcagtaac aatgcctcca gccacatagc tttgggcact tctgagtcca
      181 cctctggcca cttctttact cctccatgaa gtgacttttc caggtcactt cttgttgtat
      241 atttgtggag gatttctact agaagcgtgc agatcaccat ctgctccaca gatcaaaggc
      301 atccatctcc taagttcccc actgaaacaa gggttaatgt tggaaaattt aattcagtga
      361 atgccactca taaaaatgaa ttgcagattt taatgaacct taattttcaa aggaatttat
      421 tctatgatat ttagctaaaa ataaaaatac aataatataa tggtataatt agttttctta
      481 aattagtata ttttctgttt ggtgcctgat agagggaaaa atggcattgt tgtacttcga
      541 agacgttttt cttcattctg tcttagcagg caaacttttt tttttttaaa tagtatttac
      601 acataatgaa atggttataa aaaatctaag cttattttca acacgtaaag agatgtttct
      661 gagtgatgtt tgtgtgttgt aaatttaagc aacctttcct taactttgaa atttttcaaa
      721 ctttgtatct ttgaaagatt tatgttaaaa gctgaaaaca tcaaaattaa gaagccgaac
      781 ccccaaattt ctgattttta agatggaaag ccatctgaac ttttacagga ttccaacagg
      841 gggcagtgaa attccacaga tgaacttgat ggctgtgctg aaaaatagat gcccagaatg
      901 taaatctctg aagagaaata tgtcttaaca cagctacctc aggaaagggg gaattattgc
      961 acttaattaa agtgtattga caccatatgt ggccccacct ctcccactcc cctttttgga
     1021 ttcattaaga attaaaaaaa aaaatcagaa actagtaaag gcaatattcc tatagagcct
     1081 gtgacttaac ccttaatcat taatcctaca aaaattacac ttaaacaaaa agctagttgg
     1141 taaacagaaa cctttcaaat tagacaattt atgttcatta tctctataac tcaaaggaat
     1201 gtcattggag agctcaaacg catttggttt gtctgagttg cgctgcattc ctcattacag
     1261 gctttcctct cctagaaagc tttctagacc tctatgaaac agacattgtg gggtgtgaaa
     1321 ctgtcaaatg cagacaaaag gcgtccaagg ggcctagtgt tgcattagaa ggcacttgtt
     1381 ttccatctca aaatcgagct tgcatgtgat gaagtctgtt tttttctaaa acttggcagc
     1441 agctgaccag taaaatttgg tccttgccgg aagtgttagc tacagcggtg gcaagggtta
     1501 atattaaacc caccacagca ttttgcacaa tctcccagga gatctttcaa tggatttcca
     1561 ctgccactgg aaataatggc tcgcagctgc agcaacccat agtaacactt gatattcccg
     1621 tcgcaagcgg accagtgctt tcagtggaaa tgagaaaatc gctagtgcgg tgctgtacat
     1681 tgtgcctggg gtcaaagttc ccaagccgca acctaaaatg acctctcagc ccgacactca
     1741 gcctcggttg cttctggatg tgtctattat cgcctcattt tatgaaatgt acatagtggc
     1801 acgcgtgtat aaaggtaaaa gccagtgaag agccctaggg aacttgacat tcctggtaca
     1861 aagggaattg atttctgttt tttcacagag tagttttcag tcttcttaaa gggaatttag
     1921 ggtattcttt caaggttgtg gagctagccc gggctcgaga tctgcgatct gcatctcaat
     1981 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     2041 tccgcccatt ctccgcccca tcgctgacta atttttttta tttatgcaga ggccgaggcc
     2101 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     2161 tgcaaaaagc ttggcattcc ggtactgttg gtaaagccac catggaagac gccaaaaaca
     2221 taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc
     2281 ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata
     2341 tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta
     2401 tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc
     2461 aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg
     2521 acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt
     2581 tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc
     2641 aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt
     2701 tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg
     2761 atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag
     2821 gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt
     2881 ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt
     2941 ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata
     3001 gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc
     3061 tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat
     3121 ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg
     3181 ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat
     3241 cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc
     3301 cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa
     3361 gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag
     3421 cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata gcttactggg
     3481 acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct
     3541 atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg
     3601 caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt
     3661 tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa
     3721 caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta
     3781 ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa
     3841 agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac atgataagat
     3901 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3961 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4021 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     4081 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tctggatccg tcgaccgatg
     4141 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     4201 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     4261 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4321 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4381 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4441 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4501 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4561 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4621 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4681 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4741 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4801 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4861 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     4921 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4981 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5041 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5101 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5161 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5221 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5281 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5341 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5401 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5461 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5521 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5581 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5641 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5701 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5761 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5821 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5881 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5941 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6001 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6061 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6121 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6181 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     6241 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     6301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     6361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     6421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     6481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     6541 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     6601 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt
     6661 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc
     6721 caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat
     6781 aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta
     6841 acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc
     6901 agtgcaagtg caggtgccag aacatttctc tatcgata
//
