LOCUS       sequence-9903-ex        5753 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 5753)
  AUTHORS   Xu N, Papagiannakopoulos T, Pan G, Thomson JA, Kosik KS
  TITLE     MicroRNA-145 regulates OCT4, SOX2, and KLF4 and represses 
            pluripotency in human embryonic stem cells.
  JOURNAL   Cell. 2009 May 15. 137(4):647-58.
  PUBMED    19409607
REFERENCE   2  (bases 1 to 5753)
  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: "Cell. 2009 
            May 15. 137(4):647-58."
COMMENT     Sequence Label: P-del (pTransluc-miR-145-2)
FEATURES             Location/Qualifiers
     source          1..5753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      969..978
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             975..2627
                     /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(1041..1059)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2668..2789
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2679..2698)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2733..2752
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3085..3102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3256..3844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3336..3355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4015..4806,4807..4875))
                     /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     4638..4657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4876..4980)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5007..5462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5094..5113)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5304..5325
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5593..5641
                     /note="synthetic polyadenylation signal"
     misc_feature    5655..5746
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5695..5714
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccta aaaacccttt cagccctgaa gataaaaatt
       61 ctggcctttg agtcagggtc ttcactctgt acttctaacc ttggctccgg ggtattgccg
      121 aaaccagtcc aggcacgtca caaatgtctg acttctccca gaggcttcag aagcacaatg
      181 agcagcagag gagagccatg gagccaagca cagtctcatt taacctcccc aaaagcttgg
      241 gaagtgggtg gtgttatagc cccattttac agatgagaaa aactgaggct tatttaagca
      301 gctcacctaa agtcacatat tgattgtgct gagctgagat tgtaccctaa tctgccttca
      361 aatccatgtt tttacccatt gcatgtgatt atggaacctg ggaccgagga gcaggaggag
      421 aacattctaa attctgctcc catcttgtct ttacatctca ggtcactttt agcaaagaca
      481 gacccggaca cttgccatta atactacagg cttccttcct cctaccccct tcccccaatc
      541 ttattcatct cacctctcca gtaggtcgtg gactcatgca ttttataatt aaagaaggcc
      601 aaggctcagg ggttaagtga cttactggag gttatcagag aggaagttgc caaacccagg
      661 ctaggaactg aatggttggt ttaaatccac ccctctggca ggagactggg gaatacacat
      721 gagccgtgca gacagcagag ggcagtcctg ggggtggggg cgccagaggg tttccggtac
      781 ttttcagggc aattgaagtt ccggtcacta ctccccccca gagcaataag ccacatccgg
      841 cgacgtgtgg caccccaccc tggctgctac agatggggct ggatgcagaa gagaactcca
      901 gctggtcctt agggacacgg agatctgggt atataatgga agcttggcat tccggtactg
      961 ttggtaaagc caccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
     1021 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
     1081 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     1141 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     1201 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     1261 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     1321 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     1381 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     1441 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     1501 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     1561 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     1621 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     1681 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     1741 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     1801 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     1861 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     1921 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     1981 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     2041 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     2101 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     2161 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     2221 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     2281 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     2341 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     2401 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     2461 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     2521 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     2581 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg
     2641 gcggccggcc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     2701 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2761 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2821 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2881 aaatcgataa ggatccgtcg accccgccaa ccagatgatc tgttctcccg ccaaccagat
     2941 gatctgttct cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3001 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3061 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3661 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4981 cacatttccc cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg
     5041 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5101 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5161 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5221 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5281 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5341 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5401 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     5461 ttcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     5521 gctattacgc cagcccaagc taccatgata agtaagtaat attaaggtac gggaggtact
     5581 tggagcggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg ttttttgtgt
     5641 gaatcgatag tactaacata cgctctccat caaaacaaaa cgaaacaaaa caaactagca
     5701 aaataggctg tccccagtgc aagtgcaggt gccagaacat ttctctatcg ata
//
