LOCUS       sequence-461191-        5609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer near CALD1, chr7:134823259-134823760 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5609)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5609)
  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_CALD1
FEATURES             Location/Qualifiers
     source          1..5609
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(600..1391,1392..1460))
                     /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     1223..1242
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1461..1565)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1592..2047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1679..1698)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1889..1910
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    2178..2226
                     /note="synthetic polyadenylation signal"
     misc_feature    2240..2331
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2280..2299
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2979..3175
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3026..3161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3205..3214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3211..4863
                     /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(3277..3295)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    4904..5025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4969..4988
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5279..5296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5450..5609,1..429))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5530..5549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
       61 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
      121 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
      181 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
      241 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      301 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      361 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      421 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      481 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
      541 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
      601 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
      661 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
      721 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
      781 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
      841 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
      901 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
      961 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     1021 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     1081 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     1141 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     1201 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     1261 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     1321 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     1381 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     1441 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     1501 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     1561 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     1621 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1681 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1741 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     1801 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     1861 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1921 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1981 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     2041 acaatttgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     2101 tcttcgctat tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag
     2161 gtacttggag cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     2221 tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac
     2281 tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgatagg
     2341 taccgagctc ttacgcgtgc tagctaaggc ccgaagcact gaatgggatg aataaaatat
     2401 ctcgatattt acatggttca ttttgaccgt acccacaggg cagcaaacat ctgcgtgaaa
     2461 agtaaagaat tcagcctgtt agatgactgg ttgtgaagag ccaaacattg ggtatcatgt
     2521 catccacatg tctggttgat tgaatatgaa ttgccccctt tttatccaga tagcttcagc
     2581 aatagataaa gcatggagat ttttccacat tttacatggt gcttggtgtt gtagaattca
     2641 gcctttgtgt gttcttttcc ccccgtcttg ctattgcctg gatctcatta ggaaccatta
     2701 gtgatgtaat actaaaccta tggaatctta aacatgcttt taaaatcctg tggaataaag
     2761 atgtctccta ccaaaccaaa aatgcagtgg gaatagacaa agttacattt cttcaagaaa
     2821 aaatgactca tttcatagtt cccaaactgt tttgactgaa gcttctcaaa ataaactttc
     2881 tggaatatgg agcctttgat gtctgacttt gaaaaaaata tatcatttct caaacataca
     2941 attcaatcct tttggggtgt tcctcgagat ctgcgatctg catctcaatt agtcagcaac
     3001 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3061 tccgccccat cgctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3121 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3181 tggcattccg gtactgttgg taaagccacc atggaagacg ccaaaaacat aaagaaaggc
     3241 ccggcgccat tctatccgct ggaagatgga accgctggag agcaactgca taaggctatg
     3301 aagagatacg ccctggttcc tggaacaatt gcttttacag atgcacatat cgaggtggac
     3361 atcacttacg ctgagtactt cgaaatgtcc gttcggttgg cagaagctat gaaacgatat
     3421 gggctgaata caaatcacag aatcgtcgta tgcagtgaaa actctcttca attctttatg
     3481 ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc ccgcgaacga catttataat
     3541 gaacgtgaat tgctcaacag tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa
     3601 aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt
     3661 atcatggatt ctaaaacgga ttaccaggga tttcagtcga tgtacacgtt cgtcacatct
     3721 catctacctc ccggttttaa tgaatacgat tttgtgccag agtccttcga tagggacaag
     3781 acaattgcac tgatcatgaa ctcctctgga tctactggtc tgcctaaagg tgtcgctctg
     3841 cctcatagaa ctgcctgcgt gagattctcg catgccagag atcctatttt tggcaatcaa
     3901 atcattccgg atactgcgat tttaagtgtt gttccattcc atcacggttt tggaatgttt
     3961 actacactcg gatatttgat atgtggattt cgagtcgtct taatgtatag atttgaagaa
     4021 gagctgtttc tgaggagcct tcaggattac aagattcaaa gtgcgctgct ggtgccaacc
     4081 ctattctcct tcttcgccaa aagcactctg attgacaaat acgatttatc taatttacac
     4141 gaaattgctt ctggtggcgc tcccctctct aaggaagtcg gggaagcggt tgccaagagg
     4201 ttccatctgc caggtatcag gcaaggatat gggctcactg agactacatc agctattctg
     4261 attacacccg agggggatga taaaccgggc gcggtcggta aagttgttcc attttttgaa
     4321 gcgaaggttg tggatctgga taccgggaaa acgctgggcg ttaatcaaag aggcgaactg
     4381 tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca atccggaagc gaccaacgcc
     4441 ttgattgaca aggatggatg gctacattct ggagacatag cttactggga cgaagacgaa
     4501 cacttcttca tcgttgaccg cctgaagtct ctgattaagt acaaaggcta tcaggtggct
     4561 cccgctgaat tggaatccat cttgctccaa caccccaaca tcttcgacgc aggtgtcgca
     4621 ggtcttcccg acgatgacgc cggtgaactt cccgccgccg ttgttgtttt ggagcacgga
     4681 aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa
     4741 aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac cggaaaactc
     4801 gacgcaagaa aaatcagaga gatcctcata aaggccaaga agggcggaaa gatcgccgtg
     4861 taattctaga gtcggggcgg ccggccgctt cgagcagaca tgataagata cattgatgag
     4921 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4981 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     5041 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     5101 ctctacaaat gtggtaaaat cgataaggat ctggatccgt cgaccgatgc ccttgagagc
     5161 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     5221 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct tccgcttcct
     5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5581 cgaccctgcc gcttaccgga tacctgtcc
//
