LOCUS       sequence-462874-        6026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near SFRP1, chr8:41338016-41339065 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6026)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 6026)
  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"
COMMENT     Sequence Label: pGL3_SFRP1
FEATURES             Location/Qualifiers
     source          1..6026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(71..90)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(750..1541,1542..1610))
                     /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     1373..1392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1611..1715)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1742..2197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1829..1848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2039..2060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2328..2376
                     /note="synthetic polyadenylation signal"
     misc_feature    2390..2481
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2430..2449
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3546..3742
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3593..3728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3772..3781
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3778..5430
                     /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(3844..3862)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5471..5592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5482..5501)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5536..5555
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5846..5863)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6017..6026,1..579))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
       61 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      661 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      721 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      781 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      841 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      901 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      961 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     1021 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     1081 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1141 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1201 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1261 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1321 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1381 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1441 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1501 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1561 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1621 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1681 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1741 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1801 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1921 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2041 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2101 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2161 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     2221 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     2281 tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt
     2341 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
     2401 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
     2461 caggtgccag aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcgcttgc
     2521 ccagaagcaa ttttgtgtaa aagtatcacg cgaattattt accagagcta tgtctcttga
     2581 cagcaatcta cagacacctc tttgtagaag caatgaaatt aaaatctgtt gcattaaacc
     2641 taactcctta agcagcccca tcttcacccc atcccccagc ccccctccag atacatgagg
     2701 atggagcaca gcagcttctc ccctattctc agcttctccc ttattcctac ctgaaactga
     2761 gtggtctccc ccaaccttgg ccaccccgcc cctgcccagg ctcagccatt gagtttacat
     2821 cttttagcct aattcatttt ctagatagat ggagcaaata cctagcaaat attatttatg
     2881 gctttgtgat aaataagcct tttacagaag tttttcaagg agttctgggg aatttcaagg
     2941 ataactttca gcaagtttca aaggtcgggt caggctggga tactgagccc atagtatttg
     3001 ccgggccgtt catctgtttt gaattattgg ccttatcaag cttccgcggg ctctcaatgg
     3061 aggagctgtg ctgtagctct caggctgtat tttatttcca tcgtgggcaa cggtctttaa
     3121 tgattatttg tgctagactg acagtgggga gaaagaaaaa aggggtcttc atgaaattca
     3181 ggctcccgca gatgtgctgg gaaagatgtt cctacctgcc tggccacccc tgcccgtgtg
     3241 cccccaaccc cgcacacagg gtgatggcat ccccccgcgt gcagccaaga ggacttggcg
     3301 gcacagagtg gaggtcagaa agggcagagt tttcccaagg gtccgggaag catgggccct
     3361 tttctgatgt atgaacaaac acacatctga ttctgtgcag acttgcaaat gtagcttctt
     3421 gggagcagag accgcgtttt gttcaactcc acagcctcgg aacctaacac tctgtctgca
     3481 catagtaggt gctcaataaa tgttggttgg attgggttgg attggattgc tcgagatctg
     3541 cgatctgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3601 cctaactccg cccagttccg cccattctcc gccccatcgc tgactaattt tttttattta
     3661 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3721 tggaggccta ggcttttgca aaaagcttgg cattccggta ctgttggtaa agccaccatg
     3781 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
     3841 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
     3901 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
     3961 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
     4021 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
     4081 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
     4141 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
     4201 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
     4261 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     4321 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     4381 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     4441 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     4501 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     4561 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     4621 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     4681 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     4741 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     4801 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     4861 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     4921 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     4981 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     5041 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     5101 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     5161 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     5221 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     5281 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     5341 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     5401 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga
     5461 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5521 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5581 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     5641 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctg
     5701 gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     5761 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     5821 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5881 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5941 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6001 ccgcgttgct ggcgtttttc catagg
//
