LOCUS       sequence-274072-        3746 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 3746)
  AUTHORS   Farh KK, Grimson A, Jan C, Lewis BP, Johnston WK, Lim LP, Burge CB, 
            Bartel DP
  TITLE     The widespread impact of mammalian MicroRNAs on mRNA repression and 
            evolution.
  JOURNAL   Science. 2005 Dec 16. 310(5755):1817-21.
  PUBMED    16308420
REFERENCE   2  (bases 1 to 3746)
  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: "Science. 
            2005 Dec 16. 310(5755):1817-21."
COMMENT     Sequence Label: pIS2
FEATURES             Location/Qualifiers
     source          1..3746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      97..232
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          316..448
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     431..450
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        493..511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             521..1456
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    1528..1649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1539..1558)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1593..1612
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1782..1886
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1887..1955,1956..2747)
                     /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(2105..2124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2918..3506
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3407..3426
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(3635..3746,1..246)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(3635..3746,1..80)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3658..3678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
       61 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      121 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      181 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
      241 tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
      301 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
      361 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
      421 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
      481 ggctagagta cttaatacga ctcactatag gctagccacc atgacttcga aagtttatga
      541 tccagaacaa aggaaacgga tgataactgg tccgcagtgg tgggccagat gtaaacaaat
      601 gaatgttctt gattcattta ttaattatta tgattcagaa aaacatgcag aaaatgctgt
      661 tattttttta catggtaacg cggcctcttc ttatttatgg cgacatgttg tgccacatat
      721 tgagccagta gcgcggtgta ttataccaga ccttattggt atgggcaaat caggcaaatc
      781 tggtaatggt tcttataggt tacttgatca ttacaaatat cttactgcat ggtttgaact
      841 tcttaattta ccaaagaaga tcatttttgt cggccatgat tggggtgctt gtttggcatt
      901 tcattatagc tatgagcatc aagataagat caaagcaata gttcacgctg aaagtgtagt
      961 agatgtgatt gaatcatggg atgaatggcc tgatattgaa gaagatattg cgttgatcaa
     1021 atctgaagaa ggagaaaaaa tggttttgga gaataacttc ttcgtggaaa ccatgttgcc
     1081 atcaaaaatc atgagaaagt tagaaccaga agaatttgca gcatatcttg aaccattcaa
     1141 agagaaaggt gaagttcgtc gtccaacatt atcatggcct cgtgaaatcc cgttagtaaa
     1201 aggtggtaaa cctgacgttg tacaaattgt taggaattat aatgcttatc tacgtgcaag
     1261 tgatgattta ccaaaaatgt ttattgaatc ggacccagga ttcttttcca atgctattgt
     1321 tgaaggtgcc aagaagtttc ctaatactga atttgtcaaa gtaaaaggtc ttcatttttc
     1381 gcaagaagat gcacctgatg aaatgggaaa atatatcaaa tcgttcgttg agcgagttct
     1441 caaaaatgaa caataattct aggagctcta taccggtctc gatatcacta ctagttgttc
     1501 tagagcggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     1561 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1621 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1681 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     1741 aaatcgataa ggatccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1801 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1861 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1921 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1981 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2041 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     2101 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     2161 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2221 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2281 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2341 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2401 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2461 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2521 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2581 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2641 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2701 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2761 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2821 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2881 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2941 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3001 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3061 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3121 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3181 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3241 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3301 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3361 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3421 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3481 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3541 ccttttgctg gccttttgct cacatggctc gacagatctg cgcagcacca tggcctgaaa
     3601 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     3661 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaacc
//
