LOCUS       sequence-466700-        5548 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Enhancer near IMPG1, chr6:76224940-76225441 cloned into pGL3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5548)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5548)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGL3_IMPG1
FEATURES             Location/Qualifiers
     source          1..5548
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1014..1135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1105..1124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(1176..2828)
                     /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     2744..2762
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      2825..2834
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2864..3060)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2878..3013
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3647..3738
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(3679..3698)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    3752..3800
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3931..4386)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4089)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4280..4299
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4413..4517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4518..4586,4587..5378)
                     /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     complement(4736..4755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgctgccg
      781 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
      841 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgac
      901 ggatccagat ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc
      961 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     1021 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1081 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1141 tctgctcgaa gcggccggcc gccccgactc tagaattaca cggcgatctt tccgcccttc
     1201 ttggccttta tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc
     1261 ggtacttcgt ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg
     1321 gcgacgtaat ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg
     1381 gcggcgggaa gttcaccggc gtcatcgtcg ggaagacctg cgacacctgc gtcgaagatg
     1441 ttggggtgtt ggagcaagat ggattccaat tcagcgggag ccacctgata gcctttgtac
     1501 ttaatcagag acttcaggcg gtcaacgatg aagaagtgtt cgtcttcgtc ccagtaagct
     1561 atgtctccag aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg
     1621 tttacataac cggacataat cataggacct ctcacacaca gttcgcctct ttgattaacg
     1681 cccagcgttt tcccggtatc cagatccaca accttcgctt caaaaaatgg aacaacttta
     1741 ccgaccgcgc ccggtttatc atccccctcg ggtgtaatca gaatagctga tgtagtctca
     1801 gtgagcccat atccttgcct gatacctggc agatggaacc tcttggcaac cgcttccccg
     1861 acttccttag agaggggagc gccaccagaa gcaatttcgt gtaaattaga taaatcgtat
     1921 ttgtcaatca gagtgctttt ggcgaagaag gagaataggg ttggcaccag cagcgcactt
     1981 tgaatcttgt aatcctgaag gctcctcaga aacagctctt cttcaaatct atacattaag
     2041 acgactcgaa atccacatat caaatatccg agtgtagtaa acattccaaa accgtgatgg
     2101 aatggaacaa cacttaaaat cgcagtatcc ggaatgattt gattgccaaa aataggatct
     2161 ctggcatgcg agaatctcac gcaggcagtt ctatgaggca gagcgacacc tttaggcaga
     2221 ccagtagatc cagaggagtt catgatcagt gcaattgtct tgtccctatc gaaggactct
     2281 ggcacaaaat cgtattcatt aaaaccggga ggtagatgag atgtgacgaa cgtgtacatc
     2341 gactgaaatc cctggtaatc cgttttagaa tccatgataa taattttttg gatgattggg
     2401 agcttttttt gcacgttcaa aattttttgc aacccctttt tggaaacgaa caccacggta
     2461 ggctgcgaaa tgcccatact gttgagcaat tcacgttcat tataaatgtc gttcgcgggc
     2521 gcaactgcaa ctccgataaa taacgcgccc aacaccggca taaagaattg aagagagttt
     2581 tcactgcata cgacgattct gtgatttgta ttcagcccat atcgtttcat agcttctgcc
     2641 aaccgaacgg acatttcgaa gtactcagcg taagtgatgt ccacctcgat atgtgcatct
     2701 gtaaaagcaa ttgttccagg aaccagggcg tatctcttca tagccttatg cagttgctct
     2761 ccagcggttc catcttccag cggatagaat ggcgccgggc ctttctttat gtttttggcg
     2821 tcttccatgg tggctttacc aacagtaccg gaatgccaag ctttttgcaa aagcctaggc
     2881 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     2941 ctgcataaat aaaaaaaatt agtcagcgat ggggcggaga atgggcggaa ctgggcggag
     3001 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     3061 gatcgcagat cgcttaagat ctcaataatt ctctccctca ctcttttgca atgttttgtt
     3121 tcctcaattc acactgagac ctgcacgaag ttgtggaaac tgctctttcc cagggagatt
     3181 aacatgaatt catcttgaag tcctaactgt gttctggcta tttaatgctg tacatttata
     3241 acctttgtct tccttcttta agctgcagtt tctttgtgtt atcacagaga taggaaaact
     3301 gacacaggaa atgcataaca catgttagac tccaagacct ctctccatat gggctcagtt
     3361 ctgatttcct ctgtttccta tatggcactg agacaaatgc ctggacacag ggttttttct
     3421 tcctaaagac tcacccagaa gttctgctat taatcatcaa tccgttcctc aaaataactt
     3481 ggcatagcta aacctagcaa agtaaggaag atctgacata tggtgaagtt ctccaacagc
     3541 aattgtggga ttacaagata acagccacaa atctgtgtgc atggctattt tacatagctc
     3601 aagacttctt gaggctagca cgcgtaagag ctcggtacct atcgatagag aaatgttctg
     3661 gcacctgcac ttgcactggg gacagcctat tttgctagtt tgttttgttt cgttttgttt
     3721 tgatggagag cgtatgttag tactatcgat tcacacaaaa aaccaacaca cagatgtaat
     3781 gaaaataaag atattttatt gcggccgctc caagtacctc ccgtacctta atattactta
     3841 cttatcatgg tagcttgggc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3901 aacagttgcg cagcctgaat ggcgaatggc aaattgtaag cgttaatatt ttgttaaaat
     3961 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4021 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4081 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4141 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4201 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4261 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4321 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4381 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttc
//
