LOCUS       sequence-108763-        8082 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3.2 kb marine armor plate enhancer with a single T->G bp change 
            driving Luciferase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8082)
  AUTHORS   O'Brown NM, Summers BR, Jones FC, Brady SD, Kingsley DM
  TITLE     A recurrent regulatory change underlying altered expression and Wnt 
            response of the stickleback armor plates gene EDA.
  JOURNAL   Elife. 2015 Jan 28;4:e05290. doi: 10.7554/eLife.05290.
  PUBMED    25629660
REFERENCE   2  (bases 1 to 8082)
  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"; journalName: "Elife."; 
            date: "2015-01-28"; volume: "4:e05290. doi"; pages: " 
            10.7554/eLife.05290"
COMMENT     Sequence Label: p3.2mar(T->G)-Luc
FEATURES             Location/Qualifiers
     source          1..8082
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3259..3290
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     regulatory      3346..3355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3352..5004
                     /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(3418..3436)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5045..5166
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5056..5075)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5110..5129
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5414..5431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5585..6173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5665..5684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6344..7135,7136..7204))
                     /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     6967..6986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7205..7309)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7336..7791
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7423..7442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7633..7654
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7922..7970
                     /note="synthetic polyadenylation signal"
     misc_feature    7984..8075
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     8024..8043
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatg tggccagata ggccacatgg
       61 tgtgggagag cagtgatcgt tctttcaaac tatttcactg tttgttttgt tttagtcatc
      121 atcatcgtct cctttgtttc cccaacagtg ttgaaggcca gaaattgatc atgtctctct
      181 cccttattgt ttcaggggtt cccgataaga caaaaacaaa agaattccag gtagagcatg
      241 tcagctattg aagtaaagtc ttaccacaga cacaagcgct gataaaacag cggctgcttt
      301 gtttaaatgt gtatcatgtg ttgctcacca gggatcttct ttctttattt tctttcagcc
      361 tgcagtggtt catttgcagg gtcaggagac gaccatccaa gtgagagaag gtggggcttc
      421 cgtccgctcg gtctcttgta gttgcgccgc tgtctttgac gcaaagtcat taacgtgtca
      481 acaccctttg tcacagaacc gggcctgcaa ctgttaccgc tacacacgga cttacaccac
      541 aacgaatata taacctgcta accatattta ctttactagg gttgtgtatg attttggtta
      601 agcggaagta atcttctagc tattacacga ttagttcagt aggcttgtat tttaaatcag
      661 acgtgtcaga cttggccaga gctgaaccct cgaatgttag caggttgaag tcatcctgca
      721 tcatttgtgt gtgtgtgtgt gtgagaatca tgagatggtt tatatattcc gccagtgctg
      781 agctctggtt gtagttaaac cctgctgaga gctgttttat gttgactgtg cgcctcttgt
      841 ttgtttgttg ttgcatgctt agatctttct gaaggcatcc tcaggaactg gaagatggtg
      901 tccatccatc accgggtgtt taaaatgcac tcccgctctg gagagctaga ggtgctgctg
      961 gacggtgtct acttcatcta tagtcaggta gaagtgagta cggtccttga cgtaactctt
     1021 atgatttaca gttcggagtg aaaagaactg gcagcttctc tgtagctaag cgtaaaaacc
     1081 tgttccattg atctacccat caatgtttga cttttattat gagaatattt tcactctccc
     1141 taggtggacg tctataactc aaacccgtgt tcgctactcc gaaattccag ctgttctagt
     1201 ttttaaataa tctctttaat atcgtgtgtg gatctctttt gactacccct cttagtcggc
     1261 tgtgaattgt atgtcctcgc caggtgtact acctcaactt caccgacatt gccagctacg
     1321 aggtgatggt ggattccaac ccgttcctcc gctgcacctg cagcatcgag acgggccagc
     1381 gcaagttcaa cacgtgctac acggccgggg tcagcctgct ccgtgccggc cagaggatct
     1441 ccatacgcat agtgtatgag gacacgctca tcagcatgac caaccacacc accttcctgg
     1501 gaagcgtcag gcttggagag gctccatctg ctggacaaaa ctgataacta cacaaccacc
     1561 gggcctctac gtgaaatgcc ccccacccct ccttcccaca ctctgttcag aggaaactta
     1621 aatcctccca gtaattgaca ttgaggaacc cccccccccc cccacttgga cttggggact
     1681 tatgtcacct tcacaggtct gcattaagaa gaagtgtata gaccgcccca ctttgccctc
     1741 acgttgactc tgtgcaaaga gatcagccgt gatgagccta aaaaacaaaa aaaagggttt
     1801 ctatttctag tcaccatctg gtgtcaacac agacgacatc tctctttata accagaaagg
     1861 acactgcgct gacatatact taatatttac atattgtcaa gtgctgctga atgataatct
     1921 cgtatcgcca aaaaactcca gctattccaa atgctctatt tgcagttttt tatagttggt
     1981 cattttaatt cattgaatta tattaacagg tataagtagg tggtttaaca tgcgcttatt
     2041 agcaccatat gccactgggt tattcaaaaa aaggttcaaa gcaagtataa agatatctat
     2101 gcaaggctga gaaaagaaac attaaaactg gagttataag gtattcacag accaacagca
     2161 ctatatattt tgcacatatt tgcactacag catccctgca gaatatcaga gatatgtttt
     2221 gttacagtgc tattgtttgg tcaccgtatg aatgaatcca tgtggaaaaa tgatagttgt
     2281 ttatacgtct ctgtacatat tgtatataaa aagggatata cttttgtaaa tggagttgat
     2341 tgatggatta aaatgtttaa ttttctgtct aaatgcattt gtctgtcttg tgtttgtgta
     2401 gctatggtat tgatttaaag ggctaaacgt gtctcctatt ttggacactg ctggcacggg
     2461 ttttgacaag tgatgttctc tgtggccttt ggctccatta aaatgttgct tttactttgc
     2521 aagttttaca acacacttca aatttggctg taatcttcgt agtatctaaa ttagttccat
     2581 cttgagaggc agagagaaga tggttcctca ttttgaaact ttacatttag gtttccgggg
     2641 gttaaccaga gtgtgacctc acccatttga gtaacttcag gacatgcagt ctgcgtcttc
     2701 cttctcacga tgctttacag cgccttattg tcactagcag catttatgtt ttgtcttagt
     2761 gttttcctgg tgcacagagc cagtgtttta gaaaatgatt tgcgcaaact ccaaggggac
     2821 ataattctac aattcaatca gccacgttct gagggagtca atgggaaaat ggccaagatg
     2881 tctgaaagaa gggaggtatg tcactgatta tggaggatgt agtcatcttg gacattgtac
     2941 tactgagatt tatgccatta gtgagtgttc agatagcgct gcaataacac attttgtcct
     3001 aatgctctgt tttctctaag gacataaagc caagtacttt gccaaacagt ctgagttctc
     3061 tgaaggtttc ttccaggaga gtgaaacggc aacagggcag ctgcagaggt gaactaaaaa
     3121 tgtgtccatt agttacggtc taatcttgta gtgagaccat aaacactata tactaagtta
     3181 attgaattaa agtattatgt ttatttcctc ctcctgccag ctccaacatg gccagatagg
     3241 ccctcgagat ctagactcta gagggtatat aatggaagct cgaattccag cttggcattc
     3301 cggtactgtt ggtaaaaagc ttggcattcc ggtactgttg gtaaagccac catggaagac
     3361 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     3421 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaacaat tgcttttaca
     3481 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     3541 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     3601 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     3661 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     3721 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     3781 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     3841 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     3901 gagtccttcg atagggacaa gacaattgca ctgatcatga actcctctgg atctactggt
     3961 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     4021 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     4081 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     4141 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     4201 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     4261 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     4321 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     4381 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     4441 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     4501 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     4561 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     4621 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     4681 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     4741 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     4801 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     4861 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     4921 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     4981 aagggcggaa agatcgccgt gtaattctag agtcggggcg gccggccgct tcgagcagac
     5041 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     5101 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     5161 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     5221 gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtcgacc
     5281 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactat
     5341 cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg tgccggcagc
     5401 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5461 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5521 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5581 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5641 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5701 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5761 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5821 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5881 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5941 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6001 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6061 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6121 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6181 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6241 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6301 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6361 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6421 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6481 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6541 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6601 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6661 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6721 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6781 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6841 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6901 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6961 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7021 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7081 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7141 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7201 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7261 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7321 gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     7381 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     7441 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     7501 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     7561 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     7621 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     7681 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     7741 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt tcccattcgc
     7801 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7861 agcccaagct accatgataa gtaagtaata ttaaggtacg ggaggtactt ggagcggccg
     7921 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagt
     7981 actaacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa aataggctgt
     8041 ccccagtgca agtgcaggtg ccagaacatt tctctatcga ta
//
