LOCUS       sequence-466705-        5689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  chr6:48472983-48473484 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5689)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5689)
  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_PTCHD4
FEATURES             Location/Qualifiers
     source          1..5689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(630..1421,1422..1490))
                     /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     1253..1272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1491..1595)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1622..2077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1709..1728)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1919..1940
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2208..2256
                     /note="synthetic polyadenylation signal"
     misc_feature    2270..2361
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2310..2329
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        3089..3285
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3136..3271
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3315..3324
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3321..4973
                     /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(3387..3405)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    5014..5135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5025..5044)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5079..5098
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5389..5406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5560..5689,1..459))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5640..5659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
       61 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      121 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      181 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      241 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      301 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
      361 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
      421 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
      481 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      541 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      601 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
      661 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
      721 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
      781 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
      841 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
      901 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
      961 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     1021 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     1081 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     1141 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     1201 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     1261 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     1321 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     1381 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     1441 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     1501 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     1561 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     1621 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1681 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1741 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     1801 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     1861 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     1921 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     1981 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2041 aacgcgaatt ttaacaaaat attaacgctt acaatttgcc attcgccatt caggctgcgc
     2101 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca
     2161 tgataagtaa gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt
     2221 tattttcatt acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc
     2281 tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg
     2341 caggtgccag aacatttctc tatcgatagg taccgagctc ttacgcgtgc tagcccttgc
     2401 tcaaccttat actcagctta attaaaaccc tgctattact aattgagaaa ctatcactta
     2461 tttgttcaaa ctaccctaac tgagccctct attaatttgt tagtaagaac actagcatct
     2521 ggagtagtat ctcatagctg ttggttgcac ttagagaaga atcttatttt ttaaattttc
     2581 tctactttta ttctaggttc agggagtaca tgtgcagatt tgttacatgg gtcaattttg
     2641 tgtcactaag gtttggtgta caaatgaaag catagaaccg aatagttttt caaccgatgt
     2701 ttccctccca ccctcccatg tcaatcagtc cacagtgtct atcattccca tctttgtgtc
     2761 cacatgtatt caatgtttag ctcctgatta taagtgagaa catgtggttt ctggttttct
     2821 cttcctgcat tagtttgctt agcataatag cttccagcta cctccatgtt gcttgcaaag
     2881 gacatgattg tgttactttg tgggagaatc tttaaagtta agaaagtatt taaaatttct
     2941 tcaagattta aaattctgct atgtttcaag attaaaacat ttaagcactt tgagtatgtg
     3001 tgaatacatg tgcatatttt gatttttcat attatttcat atttcatacg tattctgaga
     3061 gtccaattgt gactcgagat ctgcgatctg catctcaatt agtcagcaac catagtcccg
     3121 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3181 cgctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3241 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tggcattccg
     3301 gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc ccggcgccat
     3361 tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg
     3421 ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg
     3481 ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata
     3541 caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg
     3601 gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat
     3661 tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc
     3721 aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt
     3781 ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc
     3841 ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac
     3901 tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa
     3961 ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg
     4021 atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg
     4081 gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa gagctgtttc
     4141 tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct
     4201 tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt
     4261 ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc
     4321 caggtatcag gcaaggatat gggctcactg agactacatc agctattctg attacacccg
     4381 agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg
     4441 tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag
     4501 gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca
     4561 aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca
     4621 tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat
     4681 tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg
     4741 acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga
     4801 cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg
     4861 gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa
     4921 aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg taattctaga
     4981 gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
     5041 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5101 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     5161 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     5221 gtggtaaaat cgataaggat ctggatccgt cgaccgatgc ccttgagagc cttcaaccca
     5281 gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc
     5341 tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga
     5401 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5461 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5521 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5581 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5641 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgtt
//
