LOCUS       sequence-348000-        5507 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 5507)
  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 5507)
  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-humanEC1.45_min1-2
FEATURES             Location/Qualifiers
     source          1..5507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      22..31
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             28..1680
                     /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(94..112)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1721..1842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1732..1751)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1786..1805
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2096..2113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2267..2855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2347..2366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3026..3817,3818..3886))
                     /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     3649..3668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3887..3991)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4018..4473
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4105..4124)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4315..4336
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4604..4652
                     /note="synthetic polyadenylation signal"
     misc_feature    4666..4757
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4706..4725
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5303..5499
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5350..5485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cattccggta ctgttggtaa agccaccatg gaagacgcca aaaacataaa gaaaggcccg
       61 gcgccattct atccgctgga agatggaacc gctggagagc aactgcataa ggctatgaag
      121 agatacgccc tggttcctgg aacaattgct tttacagatg cacatatcga ggtggacatc
      181 acttacgctg agtacttcga aatgtccgtt cggttggcag aagctatgaa acgatatggg
      241 ctgaatacaa atcacagaat cgtcgtatgc agtgaaaact ctcttcaatt ctttatgccg
      301 gtgttgggcg cgttatttat cggagttgca gttgcgcccg cgaacgacat ttataatgaa
      361 cgtgaattgc tcaacagtat gggcatttcg cagcctaccg tggtgttcgt ttccaaaaag
      421 gggttgcaaa aaattttgaa cgtgcaaaaa aagctcccaa tcatccaaaa aattattatc
      481 atggattcta aaacggatta ccagggattt cagtcgatgt acacgttcgt cacatctcat
      541 ctacctcccg gttttaatga atacgatttt gtgccagagt ccttcgatag ggacaagaca
      601 attgcactga tcatgaactc ctctggatct actggtctgc ctaaaggtgt cgctctgcct
      661 catagaactg cctgcgtgag attctcgcat gccagagatc ctatttttgg caatcaaatc
      721 attccggata ctgcgatttt aagtgttgtt ccattccatc acggttttgg aatgtttact
      781 acactcggat atttgatatg tggatttcga gtcgtcttaa tgtatagatt tgaagaagag
      841 ctgtttctga ggagccttca ggattacaag attcaaagtg cgctgctggt gccaacccta
      901 ttctccttct tcgccaaaag cactctgatt gacaaatacg atttatctaa tttacacgaa
      961 attgcttctg gtggcgctcc cctctctaag gaagtcgggg aagcggttgc caagaggttc
     1021 catctgccag gtatcaggca aggatatggg ctcactgaga ctacatcagc tattctgatt
     1081 acacccgagg gggatgataa accgggcgcg gtcggtaaag ttgttccatt ttttgaagcg
     1141 aaggttgtgg atctggatac cgggaaaacg ctgggcgtta atcaaagagg cgaactgtgt
     1201 gtgagaggtc ctatgattat gtccggttat gtaaacaatc cggaagcgac caacgccttg
     1261 attgacaagg atggatggct acattctgga gacatagctt actgggacga agacgaacac
     1321 ttcttcatcg ttgaccgcct gaagtctctg attaagtaca aaggctatca ggtggctccc
     1381 gctgaattgg aatccatctt gctccaacac cccaacatct tcgacgcagg tgtcgcaggt
     1441 cttcccgacg atgacgccgg tgaacttccc gccgccgttg ttgttttgga gcacggaaag
     1501 acgatgacgg aaaaagagat cgtggattac gtcgccagtc aagtaacaac cgcgaaaaag
     1561 ttgcgcggag gagttgtgtt tgtggacgaa gtaccgaaag gtcttaccgg aaaactcgac
     1621 gcaagaaaaa tcagagagat cctcataaag gccaagaagg gcggaaagat cgccgtgtaa
     1681 ttctagagtc ggggcggccg gccgcttcga gcagacatga taagatacat tgatgagttt
     1741 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1801 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     1861 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
     1921 tacaaatgtg gtaaaatcga taaggatctg gatccgtcga ccgatgccct tgagagcctt
     1981 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac
     2041 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc
     2101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4021 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4081 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4141 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4201 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4261 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4321 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4381 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4441 cgaattttaa caaaatatta acgcttacaa tttgccattc gccattcagg ctgcgcaact
     4501 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagcccaag ctaccatgat
     4561 aagtaagtaa tattaaggta cgggaggtac ttggagcggc cgcaataaaa tatctttatt
     4621 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gtactaacat acgctctcca
     4681 tcaaaacaaa acgaaacaaa acaaactagc aaaataggct gtccccagtg caagtgcagg
     4741 tgccagaaca tttctctatc gataggtacg ccatgtagct gctcttccgt ggacgcggat
     4801 gcttcctgcc tcatagccgg gtcgcctccg cgctaacccc gttgaacctt gcaaccagcc
     4861 cctctagagt gtggaagaaa ggagcatcga cccagcctca ggtttcactt ctgttctagt
     4921 ttatctatct aacttagacc acatgtctgt gctttcagaa aattttccca tttttgaatc
     4981 actaacagta ccctttattc ttactactgc cagggattca ctatttttaa acagctcgag
     5041 cctcagtgca aaggtttgaa atcacagctg gctgtgatta gaaggattgg gagagatcag
     5101 tgcttgatta actttcagct gctattggtt tgcctcagtt cctgcaggag aattaatctg
     5161 taggttcgag taaattttgg atcttaatgg ataccagata cattgagccc acccagacca
     5221 ccaagatttt gcaatccttt tcctcacatt taaatggtac aaatgaattc attcctcctt
     5281 actcagtgaa agatctgcga tctgcatctc aattagtcag caaccatagt cccgccccta
     5341 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatcgctga
     5401 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5461 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgg
//
