LOCUS       sequence-341707-        5282 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 5282)
  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 5282)
  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
FEATURES             Location/Qualifiers
     source          1..5282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      26..35
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             32..1684
                     /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(98..116)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    1725..1846
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1736..1755)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1790..1809
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2100..2117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2271..2859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2351..2370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3030..3821,3822..3890))
                     /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     3653..3672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3891..3995)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4022..4477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4109..4128)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4319..4340
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    4608..4656
                     /note="synthetic polyadenylation signal"
     misc_feature    4670..4761
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4710..4729
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        5082..5278
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5129..5264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ttggcattcc ggtactgttg gtaaagccac catggaagac gccaaaaaca taaagaaagg
       61 cccggcgcca ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat
      121 gaagagatac gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga
      181 catcacttac gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata
      241 tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat
      301 gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa
      361 tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa
      421 aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat
      481 tatcatggat tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc
      541 tcatctacct cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa
      601 gacaattgca ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct
      661 gcctcataga actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca
      721 aatcattccg gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt
      781 tactacactc ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga
      841 agagctgttt ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac
      901 cctattctcc ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca
      961 cgaaattgct tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag
     1021 gttccatctg ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct
     1081 gattacaccc gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga
     1141 agcgaaggtt gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact
     1201 gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc
     1261 cttgattgac aaggatggat ggctacattc tggagacata gcttactggg acgaagacga
     1321 acacttcttc atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc
     1381 tcccgctgaa ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc
     1441 aggtcttccc gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg
     1501 aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa
     1561 aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact
     1621 cgacgcaaga aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt
     1681 gtaattctag agtcggggcg gccggccgct tcgagcagac atgataagat acattgatga
     1741 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1801 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1861 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1921 cctctacaaa tgtggtaaaa tcgataagga tctggatccg tcgaccgatg cccttgagag
     1981 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     2041 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc
     2101 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2161 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2221 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2281 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2341 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2401 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2461 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2521 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2581 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2641 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2701 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2761 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2821 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2881 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2941 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3001 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3061 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3121 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3181 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3241 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3301 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3361 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3421 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3481 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3541 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3601 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3661 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3721 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3781 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3841 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3901 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3961 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4021 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4081 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4141 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4201 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4261 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4321 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     4381 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4441 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     4501 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     4561 tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt
     4621 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
     4681 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
     4741 caggtgccag aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcccgggc
     4801 gccatgtagc tgctcttccg tggacgcgga tgcttcctgc ctcatagccg ggtcgcctcc
     4861 gcgctaaccc cgttgaacct tgcaaccagc ccctctagag tgtggaagaa aggagcatcg
     4921 acccagcctc aggtttcact tctgttctag tttatctatc taacttagac cacatgtctg
     4981 tgctttcaga aaattttccc atttttgaat cactaacagt accctttatt cttactactg
     5041 ccagggattc actattttta aacagctcga gatctgcgat ctgcatctca attagtcagc
     5101 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5161 ttctccgccc catcgctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5221 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5281 gc
//
