LOCUS       sequence-233816-        4432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Renilla luciferase reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4432)
  TITLE     Petr Svoboda lab unpublished
REFERENCE   2  (bases 1 to 4432)
  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: RL-Lin28
FEATURES             Location/Qualifiers
     source          1..4432
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        114..471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        114..305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(137..157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      322..457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          541..673
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     656..675
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        718..736
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      740..749
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             746..1681
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2439..2560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2450..2469)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2504..2523
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2693..2797
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2798..2866,2867..3658)
                     /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(3016..3035)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3829..4417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4318..4337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg acagatctgc
       61 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
      121 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
      181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
      241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
      481 tcttctgaca caacagtctc gaacttaagc tgcagaagtt ggtcgtgagg cactgggcag
      541 gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg cttgtcgaga
      601 cagagaagac tcttgcgttt ctgataggca cctattggtc ttactgacat ccactttgcc
      661 tttctctcca caggtgtcca ctcccagttc aattacagct cttaaggcta gagtacttaa
      721 tacgactcac tataggctag ccaccatggc ttccaaggtg tacgaccccg agcaacgcaa
      781 acgcatgatc actgggcctc agtggtgggc tcgctgcaag caaatgaacg tgctggactc
      841 cttcatcaac tactatgatt ccgagaagca cgccgagaac gccgtgattt ttctgcatgg
      901 taacgctgcc tccagctacc tgtggaggca cgtcgtgcct cacatcgagc ccgtggctag
      961 atgcatcatc cctgatctga tcggaatggg taagtccggc aagagcggga atggctcata
     1021 tcgcctcctg gatcactaca agtacctcac cgcttggttc gagctgctga accttccaaa
     1081 gaaaatcatc tttgtgggcc acgactgggg ggcttgtctg gcctttcact actcctacga
     1141 gcaccaagac aagatcaagg ccatcgtcca tgctgagagt gtcgtggacg tgatcgagtc
     1201 ctgggacgag tggcctgaca tcgaggagga tatcgccctg atcaagagcg aagagggcga
     1261 gaaaatggtg cttgagaata acttcttcgt cgagaccatg ctcccaagca agatcatgcg
     1321 gaaactggag cctgaggagt tcgctgccta cctggagcca ttcaaggaga agggcgaggt
     1381 tagacggcct accctctcct ggcctcgcga gatccctctc gttaagggag gcaagcccga
     1441 cgtcgtccag attgtccgca actacaacgc ctaccttcgg gccagcgacg atctgcctaa
     1501 gatgttcatc gagtccgacc ctgggttctt ttccaacgct attgtcgagg gagctaagaa
     1561 gttccctaac accgagttcg tgaaggtgaa gggcctccac ttcagccagg aggacgctcc
     1621 agatgaaatg ggtaagtaca tcaagagctt cgtggagcgc gtgctgaaga acgagcagta
     1681 attctagaaa gatgatctag agtgcctgta ctcttatgtg aaatcctcac acatacatac
     1741 agattcaagt ctcccctctc acttatgcac atactatatg tagtatctca attctataaa
     1801 acatgacacc ctgcttaaat gtagtttgct taagagtgat ttgtctaatt aagaacttcc
     1861 cagttcctca gcagtagtta acatcccggc cggctgctct ctgtggacca ctattcccct
     1921 gtgcttgcat gaggtagact tcccgcccgg gtgaagggaa cactgacaag tactaagtct
     1981 ttttccgttt ctgaatcaaa ggaagcttgg tttgacactt gtttcgcttc ccgtctggca
     2041 agggaaatat cacacagcct tctacaaaag aacttcccta gcccagccac ttggcaaagc
     2101 ctcgggttct agggtggttc ctccccacaa aagccttcac ttgctttcag atagagtgtg
     2161 gtaggagttc ctttgtctca tcctttcctt ccctccaagg aaagacaggg tgacactggg
     2221 aaccccggcc tgagacacgg cagtgcaagc tacccacgca acccaccctt acccccactt
     2281 tctccactct gcagattgat gcctctttcc ttaaactccc cattagccaa agaataaccc
     2341 tgactcctgg gcctcaattc tgggcttctg ggagcagggc agggctgtgg atctcttcct
     2401 cttcctccct ctagagcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     2461 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2521 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2581 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     2641 caaatgtggt aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac
     2701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2761 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2821 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2881 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2941 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3001 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3061 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3121 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3181 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3241 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3301 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3361 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3421 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3481 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3541 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3601 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3661 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3721 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3781 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3841 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3901 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3961 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4021 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4081 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4141 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4201 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4261 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4321 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4381 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cg
//
