LOCUS       sequence-7202-ex        4379 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 4379)
  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 4379)
  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: pAG81 ZAP3 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    694..815
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(705..724)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     759..778
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        948..1052
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1053..1121,1122..1913)
                     /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(1271..1290)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2084..2672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2573..2592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2801..3158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2801..2992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2824..2844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3009..3144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3228..3360
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3343..3362
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3405..3423
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3433..4368
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 caatcaatta catatgaaag ggcattctta gaatgcagca gatatgtggc acagcctacc
       61 caacatatgc ataattgtct cttctttgct acaagatctc caaatttgag tttctgtcaa
      121 cagaggcaga agaacttatt gaatggggag gcagagggtt gccactggct gctagatttg
      181 ttcttctgca ttttatccta agtgacgggg aaaaattaaa tgactgtgta ggcaatggat
      241 tttgctcagg ctgattagag gcttacatag tgtgagttta aagtttctgt gtacctaatg
      301 taaaatgctt tcgatcacac ctctgtagag atgtttgcct ttccctgtca ttcaagaata
      361 tgcgacctat agagttacgt tgacttttca gtgtttactg aagtacagag attagcaatg
      421 ccttggaata gtgttgcttt ttaaatccag aatattactc ggtttattta atacttgagc
      481 ccttgctaaa gttgctcctc atattctttt gaaaatgtac aaaatgagaa ctctgattcc
      541 taccccctag agtatacaga atctccaggg agtggtccca tcgaacccgt gtgtgatata
      601 gaggactctt gataaaaact gtgtttatat aaagatggtg tcaacctcta gaatgcaaga
      661 tttctctaga gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa
      721 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      781 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
      841 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
      901 gtggtaaaat cgataaggat ccaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      961 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1021 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1081 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1141 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1201 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1261 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1321 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1381 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1441 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1501 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1561 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1621 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1681 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1741 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1801 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1861 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1921 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1981 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2041 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2101 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2161 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2221 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2281 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2341 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2401 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2461 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2521 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2581 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2641 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2701 tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatctgcgca gcaccatggc
     2761 ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc
     2821 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2881 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2941 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3001 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3061 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     3121 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgattct tctgacacaa
     3181 cagtctcgaa cttaagctgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg
     3241 ttacaagaca ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct
     3301 tgcgtttctg ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag
     3361 gtgtccactc ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat
     3421 aggctagcca ccatgacttc gaaagtttat gatccagaac aaaggaaacg gatgataact
     3481 ggtccgcagt ggtgggccag atgtaaacaa atgaatgttc ttgattcatt tattaattat
     3541 tatgattcag aaaaacatgc agaaaatgct gttatttttt tacatggtaa cgcggcctct
     3601 tcttatttat ggcgacatgt tgtgccacat attgagccag tagcgcggtg tattatacca
     3661 gaccttattg gtatgggcaa atcaggcaaa tctggtaatg gttcttatag gttacttgat
     3721 cattacaaat atcttactgc atggtttgaa cttcttaatt taccaaagaa gatcattttt
     3781 gtcggccatg attggggtgc ttgtttggca tttcattata gctatgagca tcaagataag
     3841 atcaaagcaa tagttcacgc tgaaagtgta gtagatgtga ttgaatcatg ggatgaatgg
     3901 cctgatattg aagaagatat tgcgttgatc aaatctgaag aaggagaaaa aatggttttg
     3961 gagaataact tcttcgtgga aaccatgttg ccatcaaaaa tcatgagaaa gttagaacca
     4021 gaagaatttg cagcatatct tgaaccattc aaagagaaag gtgaagttcg tcgtccaaca
     4081 ttatcatggc ctcgtgaaat cccgttagta aaaggtggta aacctgacgt tgtacaaatt
     4141 gttaggaatt ataatgctta tctacgtgca agtgatgatt taccaaaaat gtttattgaa
     4201 tcggacccag gattcttttc caatgctatt gttgaaggtg ccaagaagtt tcctaatact
     4261 gaatttgtca aagtaaaagg tcttcatttt tcgcaagaag atgcacctga tgaaatggga
     4321 aaatatatca aatcgttcgt tgagcgagtt ctcaaaaatg aacaataatt ctaggagct
//
