LOCUS       sequence-354661-        5563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5563)
  AUTHORS   Renganathan A, Kresoja-Rakic J, Echeverry N, Ziltener G, Vrugt B, 
            Opitz I, Stahel RA, Felley-Bosco E
  TITLE     GAS5 long non-coding RNA in malignant pleural mesothelioma.
  JOURNAL   Mol Cancer. 2014 May 23;13:119. doi: 10.1186/1476-4598-13-119.
  PUBMED    24885398
REFERENCE   2  (bases 1 to 5563)
  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: "Mol 
            Cancer."; date: "2014-05-23"; volume: "13:119. doi"; pages: " 
            10.1186/1476-4598-13-119"
COMMENT     Sequence Label: pGL3-B-pGAS5
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      827..836
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             833..2485
                     /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(899..917)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2526..2647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2537..2556)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2591..2610
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2895..2912)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3066..3654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3146..3165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3825..4616,4617..4685))
                     /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     4448..4467
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4686..4790)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4817..5272
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4904..4923)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5114..5135
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5403..5451
                     /note="synthetic polyadenylation signal"
     misc_feature    5465..5556
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5505..5524
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagctcag gtgagaacta ggaagggggc ggcgcccagg
       61 gaccggctcc aggcggcggc gggcggggag ctgccaggag gtgggtgtgc gtatctgggt
      121 gtgtcccggc tgcggtgggg gagggggccg cccttcccgg ggtcctgagg gaggggcccc
      181 gcgagggggc gggtccggca gatcagcccg gccgggggcc aagggggtgg aaggtcgccc
      241 ggtgcctgga ggcggggtga aggaaagtag ctgagcgccc caggagcgcg ctgcggcgga
      301 gggcgggcga ggtgaccccg gcgcagggaa gtggcactgg ggtgtggggc ccgagtccag
      361 tcaaggagca gcacctacct ccgggggtgg aaatgggccg gaatgggccg gaacaccgtc
      421 ccggaagtga aatcccccac cccctcccac ggagcgggcg acgtgccgga aggaaatcac
      481 tcagccttac accgcccccc cttcccccat ccccagagct ttccttgcct cgcgccccct
      541 cccctctctg ctcttcctcc tcagtcggga ggagggcggg agcacggcat cacgtggacg
      601 gtcatgtctc tgcccacaat ggcgaggaag ggcggggaaa aggggcggac ttaaggggtg
      661 ggaagaaggt gatactggat gagaggaaga ttgagggggg ttagggggag gagccgaagc
      721 ggacgctgcc gggagcgtgc gtcgtgacgt catcaaaaga actcttatat acaggagccc
      781 aggcaccata ctgtcttaag cttggcattc cggtactgtt ggtaaagcca ccatggaaga
      841 cgccaaaaac ataaagaaag gcccggcgcc attctatccg ctggaagatg gaaccgctgg
      901 agagcaactg cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac
      961 agatgcacat atcgaggtgg acatcactta cgctgagtac ttcgaaatgt ccgttcggtt
     1021 ggcagaagct atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga
     1081 aaactctctt caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc
     1141 gcccgcgaac gacatttata atgaacgtga attgctcaac agtatgggca tttcgcagcc
     1201 taccgtggtg ttcgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaagct
     1261 cccaatcatc caaaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc
     1321 gatgtacacg ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtgcc
     1381 agagtccttc gatagggaca agacaattgc actgatcatg aactcctctg gatctactgg
     1441 tctgcctaaa ggtgtcgctc tgcctcatag aactgcctgc gtgagattct cgcatgccag
     1501 agatcctatt tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt
     1561 ccatcacggt tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt
     1621 cttaatgtat agatttgaag aagagctgtt tctgaggagc cttcaggatt acaagattca
     1681 aagtgcgctg ctggtgccaa ccctattctc cttcttcgcc aaaagcactc tgattgacaa
     1741 atacgattta tctaatttac acgaaattgc ttctggtggc gctcccctct ctaaggaagt
     1801 cggggaagcg gttgccaaga ggttccatct gccaggtatc aggcaaggat atgggctcac
     1861 tgagactaca tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg
     1921 taaagttgtt ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg
     1981 cgttaatcaa agaggcgaac tgtgtgtgag aggtcctatg attatgtccg gttatgtaaa
     2041 caatccggaa gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat
     2101 agcttactgg gacgaagacg aacacttctt catcgttgac cgcctgaagt ctctgattaa
     2161 gtacaaaggc tatcaggtgg ctcccgctga attggaatcc atcttgctcc aacaccccaa
     2221 catcttcgac gcaggtgtcg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc
     2281 cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc
     2341 cagtcaagta acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc
     2401 gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa
     2461 gaagggcgga aagatcgccg tgtaattcta gagtcggggc ggccggccgc ttcgagcaga
     2521 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     2581 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2641 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2701 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgtcgac
     2761 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     2821 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     2881 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2941 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3001 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3061 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3121 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3181 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3241 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3301 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3361 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3421 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3481 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3541 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3601 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3661 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3721 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3781 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3841 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3901 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3961 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4021 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4081 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4141 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4201 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4261 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4321 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4381 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4441 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4501 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4561 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4621 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4681 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4741 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4801 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4861 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4921 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4981 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5041 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5101 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5161 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5221 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg
     5281 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     5341 cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact tggagcggcc
     5401 gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt gaatcgatag
     5461 tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca aaataggctg
     5521 tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
