LOCUS       sequence-341637-        5735 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 5735)
  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 5735)
  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_S2
FEATURES             Location/Qualifiers
     source          1..5735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        767..963
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      814..949
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      993..1002
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             999..2651
                     /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(1065..1083)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2692..2813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2757..2776
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3067..3084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3238..3826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3318..3337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3997..4788,4789..4857))
                     /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     4620..4639
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4858..4962)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4989..5444
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5076..5095)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5286..5307
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5575..5623
                     /note="synthetic polyadenylation signal"
     misc_feature    5637..5728
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5677..5696
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcaaacgcat ttggtttgtc tgagttgcgc tgcattcctc attacaggct
       61 ttcctctcct agaaagcttt ctagacctct atgaaacaga cattgtgggg tgtgaaactg
      121 tcaaatgcag acaaaaggcg tccaaggggc ctagtgttgc attagaaggc acttgttttc
      181 catctcaaaa tcgagcttgc atgtgatgaa gtctgttttt ttctaaaact tggcagcagc
      241 tgaccagtaa aatttggtcc ttgccggaag tgttagctac agcggtggca agggttaata
      301 ttaaacccac cacagcattt tgcacaatct cccaggagat ctttcaatgg atttccactg
      361 ccactggaaa taatggctcg cagctgcagc aacccatagt aacacttgat attcccgtcg
      421 caagcggacc agtgctttca gtggaaatga gaaaatcgct agtgcggtgc tgtacattgt
      481 gcctggggtc aaagttccca agccgcaacc taaaatgacc tctcagcccg acactcagcc
      541 tcggttgctt ctggatgtgt ctattatcgc ctcattttat gaaatgtaca tagtggcacg
      601 cgtgtataaa ggtaaaagcc agtgaagagc cctagggaac ttgacattcc tggtacaaag
      661 ggaattgatt tctgtttttt cacagagtag ttttcagtct tcttaaaggg aatttagggt
      721 attctttcaa ggttgtggag ctagcccggg ctcgagatct gcgatctgca tctcaattag
      781 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
      841 gcccattctc cgccccatcg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
      901 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
      961 aaaaagcttg gcattccggt actgttggta aagccaccat ggaagacgcc aaaaacataa
     1021 agaaaggccc ggcgccattc tatccgctgg aagatggaac cgctggagag caactgcata
     1081 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
     1141 aggtggacat cacttacgct gagtacttcg aaatgtccgt tcggttggca gaagctatga
     1201 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     1261 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1321 tttataatga acgtgaattg ctcaacagta tgggcatttc gcagcctacc gtggtgttcg
     1381 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaagctccca atcatccaaa
     1441 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1501 tcacatctca tctacctccc ggttttaatg aatacgattt tgtgccagag tccttcgata
     1561 gggacaagac aattgcactg atcatgaact cctctggatc tactggtctg cctaaaggtg
     1621 tcgctctgcc tcatagaact gcctgcgtga gattctcgca tgccagagat cctatttttg
     1681 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     1741 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     1801 ttgaagaaga gctgtttctg aggagccttc aggattacaa gattcaaagt gcgctgctgg
     1861 tgccaaccct attctccttc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     1921 atttacacga aattgcttct ggtggcgctc ccctctctaa ggaagtcggg gaagcggttg
     1981 ccaagaggtt ccatctgcca ggtatcaggc aaggatatgg gctcactgag actacatcag
     2041 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     2101 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcaaagag
     2161 gcgaactgtg tgtgagaggt cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     2221 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     2281 aagacgaaca cttcttcatc gttgaccgcc tgaagtctct gattaagtac aaaggctatc
     2341 aggtggctcc cgctgaattg gaatccatct tgctccaaca ccccaacatc ttcgacgcag
     2401 gtgtcgcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2461 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2521 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     2581 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaaga
     2641 tcgccgtgta attctagagt cggggcggcc ggccgcttcg agcagacatg ataagataca
     2701 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2761 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2821 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     2881 agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct ggatccgtcg accgatgccc
     2941 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     3001 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc
     3061 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3121 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3181 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3241 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3301 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3361 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3421 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3481 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3541 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3601 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3661 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3721 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3781 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3841 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3901 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3961 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4021 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4081 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4141 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4201 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4261 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4321 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4381 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4441 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4501 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4561 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4621 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4681 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4741 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4801 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4861 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4921 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4981 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5041 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     5101 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     5161 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     5221 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     5281 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     5341 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5401 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag
     5461 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa
     5521 gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa
     5581 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
     5641 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
     5701 gcaagtgcag gtgccagaac atttctctat cgata
//
