LOCUS       sequence-297063-        3705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3705)
  AUTHORS   Yadav L, Pietila E, Ohman T, Liu X, Mahato AK, Sidorova Y, Lehti K, 
            Saarma M, Varjosalo M
  TITLE     PTPRA Phosphatase Regulates GDNF-Dependent RET Signaling and 
            Inhibits the RET Mutant MEN2A Oncogenic Potential.
  JOURNAL   iScience. 2020 Feb 21;23(2):100871. doi: 10.1016/j.isci.2020.100871.
            Epub 2020 Jan 31.
  PUBMED    32062451
REFERENCE   2  (bases 1 to 3705)
  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"; journalName: "iScience.";
            date: "2020-02-21"; volume: "23(2):100871. doi"; pages: " 
            10.1016/j.isci.2020.100871. Epub 2020 Jan 31"
COMMENT     Sequence Label: phR-Luc
FEATURES             Location/Qualifiers
     source          1..3705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..358
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1..192
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(24..44)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      209..344
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          428..560
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     543..562
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        605..623
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             633..1568
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1599..1720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1610..1629)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1664..1683
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1853..1957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1958..2026,2027..2818)
                     /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(2176..2195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2989..3577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3478..3497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
       61 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
      241 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
      301 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
      361 gcttgattct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      421 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      481 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      541 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      601 tacttaatac gactcactat aggctagcca ccatgacttc gaaagtttat gatccagaac
      661 aaaggaaacg gatgataact ggtccgcagt ggtgggccag atgtaaacaa atgaatgttc
      721 ttgattcatt tattaattat tatgattcag aaaaacatgc agaaaatgct gttatttttt
      781 tacatggtaa cgcggcctct tcttatttat ggcgacatgt tgtgccacat attgagccag
      841 tagcgcggtg tattatacca gaccttattg gtatgggcaa atcaggcaaa tctggtaatg
      901 gttcttatag gttacttgat cattacaaat atcttactgc atggtttgaa cttcttaatt
      961 taccaaagaa gatcattttt gtcggccatg attggggtgc ttgtttggca tttcattata
     1021 gctatgagca tcaagataag atcaaagcaa tagttcacgc tgaaagtgta gtagatgtga
     1081 ttgaatcatg ggatgaatgg cctgatattg aagaagatat tgcgttgatc aaatctgaag
     1141 aaggagaaaa aatggttttg gagaataact tcttcgtgga aaccatgttg ccatcaaaaa
     1201 tcatgagaaa gttagaacca gaagaatttg cagcatatct tgaaccattc aaagagaaag
     1261 gtgaagttcg tcgtccaaca ttatcatggc ctcgtgaaat cccgttagta aaaggtggta
     1321 aacctgacgt tgtacaaatt gttaggaatt ataatgctta tctacgtgca agtgatgatt
     1381 taccaaaaat gtttattgaa tcggacccag gattcttttc caatgctatt gttgaaggtg
     1441 ccaagaagtt tcctaatact gaatttgtca aagtaaaagg tcttcatttt tcgcaagaag
     1501 atgcacctga tgaaatggga aaatatatca aatcgttcgt tgagcgagtt ctcaaaaatg
     1561 aacaataatt ctagagcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     1621 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1681 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1741 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1801 caaatgtggt aaaatcgata aggatccagg tggcactttt cggggaaatg tgcgcggaac
     1861 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     1921 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     1981 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2041 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2101 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2161 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2221 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2281 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2341 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2401 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2461 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     2521 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     2581 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     2641 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     2701 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     2761 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     2821 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     2881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     2941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3601 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct gcgcagcacc
     3661 atggcctgaa ataacctctg aaagaggaac ttggttaggt acctt
//
