LOCUS       sequence-7206-ex        4536 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 4536)
  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 4536)
  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: pAG73 PFTK1 3'UTR mut
FEATURES             Location/Qualifiers
     source          1..4536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    856..977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(867..886)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     921..940
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1110..1214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1215..1283,1284..2075)
                     /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(1433..1452)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2246..2834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2735..2754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2963..3320
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2963..3154
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2986..3006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3171..3306
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          3390..3522
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     3505..3524
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        3567..3585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3595..4530
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 gagctcaaaa aatacatttt aaaaaaagaa tgcaagccaa ttggaagaca tcattgggtt
       61 cttactttaa gacatctcct ggaataactg ttcaaatgca ggttttagaa acaatgcagg
      121 aatcttgctt taaagatgaa aaagggaatg ggccagcttc ccttactcaa ggagttgagg
      181 gaccttggag gatgaaggcg agtatgtgac actggagaaa agtggaccag gcatgtcttt
      241 tgctttgatc tggagggagg gctgcctgat gcaggccggc tcccagtggg gcaggcctcg
      301 ctgcagaatg cccagtagta ctgcggccaa ggggacagtt aggagacttc atctaaagca
      361 tgaaacctag ctcctctaca cacaaattcc tatggaaata cctttgtgta cagtgtctta
      421 cattttccta ttagtcagaa agaaggagag aatgagtgag tgcttgaaat gtgtcatact
      481 gttttaggat caagactagg aattaggagc caggttgaca aggacttttt ctgagagttg
      541 ggtgagggta aagcttttct ataatcaagc tcaatacacc aaggaaactg gatccagaat
      601 tcctaaactt taaaatggta ctgtctgcgg agtggagtat ggatggttat gtcaaagtca
      661 tagttcatcc tatccagatg tagcattcat ggtaaacttt taagtgctaa gcaaggaatt
      721 atttactgat tggttttaaa gagagcagaa aacacccaag tgtagaatgt ctactgtttg
      781 ctacctagaa atcttttcca ttcctctttc atagaatgca acccactcta gagcggccgc
      841 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
      901 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
      961 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     1021 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg
     1081 atccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1141 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1201 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1261 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1321 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1381 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1441 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1501 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1561 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     1621 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     1681 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     1741 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     1801 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     1861 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     1921 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     1981 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2041 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2101 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2161 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2221 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2281 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2341 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     2401 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2461 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2521 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     2581 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     2641 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     2701 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     2761 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2821 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2881 cttttgctca catggctcga cagatctgcg cagcaccatg gcctgaaata acctctgaaa
     2941 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3001 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3061 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3121 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3181 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3241 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3301 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaagct
     3361 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
     3421 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
     3481 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
     3541 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc caccatgact
     3601 tcgaaagttt atgatccaga acaaaggaaa cggatgataa ctggtccgca gtggtgggcc
     3661 agatgtaaac aaatgaatgt tcttgattca tttattaatt attatgattc agaaaaacat
     3721 gcagaaaatg ctgttatttt tttacatggt aacgcggcct cttcttattt atggcgacat
     3781 gttgtgccac atattgagcc agtagcgcgg tgtattatac cagaccttat tggtatgggc
     3841 aaatcaggca aatctggtaa tggttcttat aggttacttg atcattacaa atatcttact
     3901 gcatggtttg aacttcttaa tttaccaaag aagatcattt ttgtcggcca tgattggggt
     3961 gcttgtttgg catttcatta tagctatgag catcaagata agatcaaagc aatagttcac
     4021 gctgaaagtg tagtagatgt gattgaatca tgggatgaat ggcctgatat tgaagaagat
     4081 attgcgttga tcaaatctga agaaggagaa aaaatggttt tggagaataa cttcttcgtg
     4141 gaaaccatgt tgccatcaaa aatcatgaga aagttagaac cagaagaatt tgcagcatat
     4201 cttgaaccat tcaaagagaa aggtgaagtt cgtcgtccaa cattatcatg gcctcgtgaa
     4261 atcccgttag taaaaggtgg taaacctgac gttgtacaaa ttgttaggaa ttataatgct
     4321 tatctacgtg caagtgatga tttaccaaaa atgtttattg aatcggaccc aggattcttt
     4381 tccaatgcta ttgttgaagg tgccaagaag tttcctaata ctgaatttgt caaagtaaaa
     4441 ggtcttcatt tttcgcaaga agatgcacct gatgaaatgg gaaaatatat caaatcgttc
     4501 gttgagcgag ttctcaaaaa tgaacaataa ttctag
//
