LOCUS       sequence-462836-        5542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IMPG1 enhancer, two sensitizing elements scrambled.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5542)
  TITLE     Wysocka Lab enhancer plasmids 2025
REFERENCE   2  (bases 1 to 5542)
  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_IMPG1_lpos0pos1scr
FEATURES             Location/Qualifiers
     source          1..5542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    565..656
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(597..616)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    670..718
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(849..1304)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(986..1007)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1198..1217
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1331..1435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1436..1504,1505..2296)
                     /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(1654..1673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2467..3055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2956..2975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3209..3226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3480..3601
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3517..3536)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3571..3590
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3642..5294)
                     /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     5210..5228
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     regulatory      5291..5300
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5330..5526)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5344..5479
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 caataattct ctccctcact cttttgcaat gttttgtttc ctcaattcac actgagacct
       61 gcacgaagtt gtggaaactg ctctttccca gggagattaa catgaattca tcttgaagtc
      121 ctaactgtgt tctggctatt taatgctgta catttataac ctttgtcttc cttctttaag
      181 ctgcagtttc tttgtgttat cacagagata ggaaaactga cacaggaaat gcataacaca
      241 tgttagactc caagacctct ctccatatgg gctcagttct gatttcctct gtttcctata
      301 tggcactgag acaaatgcct ggacacaggg ttttttcttc ctaaagactc acccagaagt
      361 tgatagtctc atcatcaatc cgttcctcaa aataacttgg catagctaaa cctagcaaag
      421 taaggaagat ctgtagtaag ctgaagttct ccaacagcaa ttgtgggatt acaagataac
      481 agccacaaat ctgtgtgcat ggctatttta catagctcaa gacttcttga ggctagcacg
      541 cgtaagagct cggtacctat cgatagagaa atgttctggc acctgcactt gcactgggga
      601 cagcctattt tgctagtttg ttttgtttcg ttttgttttg atggagagcg tatgttagta
      661 ctatcgattc acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc
      721 ggccgctcca agtacctccc gtaccttaat attacttact tatcatggta gcttgggctg
      781 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
      841 cgaatggcaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
      901 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
      961 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     1021 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     1081 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     1141 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     1201 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     1261 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     1321 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1381 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1441 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1501 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1561 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1621 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     1681 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1741 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1801 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1861 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1921 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     1981 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2041 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2101 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2161 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2221 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2281 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     2641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3181 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3241 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     3301 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     3361 gtcgacggat ccagatcctt atcgatttta ccacatttgt agaggtttta cttgctttaa
     3421 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3481 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3541 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3601 atcatgtctg ctcgaagcgg ccggccgccc cgactctaga attacacggc gatctttccg
     3661 cccttcttgg cctttatgag gatctctctg atttttcttg cgtcgagttt tccggtaaga
     3721 cctttcggta cttcgtccac aaacacaact cctccgcgca actttttcgc ggttgttact
     3781 tgactggcga cgtaatccac gatctctttt tccgtcatcg tctttccgtg ctccaaaaca
     3841 acaacggcgg cgggaagttc accggcgtca tcgtcgggaa gacctgcgac acctgcgtcg
     3901 aagatgttgg ggtgttggag caagatggat tccaattcag cgggagccac ctgatagcct
     3961 ttgtacttaa tcagagactt caggcggtca acgatgaaga agtgttcgtc ttcgtcccag
     4021 taagctatgt ctccagaatg tagccatcca tccttgtcaa tcaaggcgtt ggtcgcttcc
     4081 ggattgttta cataaccgga cataatcata ggacctctca cacacagttc gcctctttga
     4141 ttaacgccca gcgttttccc ggtatccaga tccacaacct tcgcttcaaa aaatggaaca
     4201 actttaccga ccgcgcccgg tttatcatcc ccctcgggtg taatcagaat agctgatgta
     4261 gtctcagtga gcccatatcc ttgcctgata cctggcagat ggaacctctt ggcaaccgct
     4321 tccccgactt ccttagagag gggagcgcca ccagaagcaa tttcgtgtaa attagataaa
     4381 tcgtatttgt caatcagagt gcttttggcg aagaaggaga atagggttgg caccagcagc
     4441 gcactttgaa tcttgtaatc ctgaaggctc ctcagaaaca gctcttcttc aaatctatac
     4501 attaagacga ctcgaaatcc acatatcaaa tatccgagtg tagtaaacat tccaaaaccg
     4561 tgatggaatg gaacaacact taaaatcgca gtatccggaa tgatttgatt gccaaaaata
     4621 ggatctctgg catgcgagaa tctcacgcag gcagttctat gaggcagagc gacaccttta
     4681 ggcagaccag tagatccaga ggagttcatg atcagtgcaa ttgtcttgtc cctatcgaag
     4741 gactctggca caaaatcgta ttcattaaaa ccgggaggta gatgagatgt gacgaacgtg
     4801 tacatcgact gaaatccctg gtaatccgtt ttagaatcca tgataataat tttttggatg
     4861 attgggagct ttttttgcac gttcaaaatt ttttgcaacc cctttttgga aacgaacacc
     4921 acggtaggct gcgaaatgcc catactgttg agcaattcac gttcattata aatgtcgttc
     4981 gcgggcgcaa ctgcaactcc gataaataac gcgcccaaca ccggcataaa gaattgaaga
     5041 gagttttcac tgcatacgac gattctgtga tttgtattca gcccatatcg tttcatagct
     5101 tctgccaacc gaacggacat ttcgaagtac tcagcgtaag tgatgtccac ctcgatatgt
     5161 gcatctgtaa aagcaattgt tccaggaacc agggcgtatc tcttcatagc cttatgcagt
     5221 tgctctccag cggttccatc ttccagcgga tagaatggcg ccgggccttt ctttatgttt
     5281 ttggcgtctt ccatggtggc tttaccaaca gtaccggaat gccaagcttt ttgcaaaagc
     5341 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5401 cggcctctgc ataaataaaa aaaattagtc agcgatgggg cggagaatgg gcggaactgg
     5461 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5521 gatgcagatc gcagatctcg ag
//
