LOCUS       sequence-9899-ex        6378 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 6378)
  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 6378)
  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-miR-145 (pTransluc-miR-145-1)
FEATURES             Location/Qualifiers
     source          1..6378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1594..1603
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1600..3252
                     /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(1666..1684)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    3293..3414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3304..3323)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3358..3377
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3710..3727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3881..4469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3961..3980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4640..5431,5432..5500))
                     /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     5263..5282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5501..5605)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5632..6087
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5719..5738)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5929..5950
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    6218..6266
                     /note="synthetic polyadenylation signal"
     misc_feature    6280..6371
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     6320..6339
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagccata ctatcagcca cttgtgatgc tggggaagtt
       61 cctctacaca agttcccctg gtgccacgat ctgcttcacg agtctgggca tgtcctgact
      121 cctctgtgta ccccagtgtg tccatcttag catgaggcgt tagcatttcc ccagcaccct
      181 gcctctgctt tctctttctg agcccatggc agacacagct aatcagtaaa ggcatatatt
      241 cttttttttt tgagacacag tctcactcca tcacccaggc tggagtgcag tggcataatc
      301 tcagctcact gcaacctcct cctcccaggg tcaagcaatt ctcgtgcctc agcctcccaa
      361 gtagctgaga ttacaggtgt gcgccaccac acctggctaa tttttgtatt tttagtagag
      421 atggggtttc actatgttgg ccaggctggt ctcaaactcc tgacttcagg tgatctgccc
      481 gcctcggcct cccaaagtgc tgggattaca ggcatgagcc tccacgcccc gccaggcatc
      541 cattcttgaa gtgctcagat gcagcttcag aatcctcaac aatgagccct ggaaagccac
      601 tagtactgat tcacagctga aactcattca tcatttgtgg atcaaacaat tcctaaaaac
      661 cctttcagcc ctgaagataa aaattctggc ctttgagtca gggtcttcac tctgtacttc
      721 taaccttggc tccggggtat tgccgaaacc agtccaggca cgtcacaaat gtctgacttc
      781 tcccagaggc ttcagaagca caatgagcag cagaggagag ccatggagcc aagcacagtc
      841 tcatttaacc tccccaaaag cttgggaagt gggtggtgtt atagccccat tttacagatg
      901 agaaaaactg aggcttattt aagcagctca cctaaagtca catattgatt gtgctgagct
      961 gagattgtac cctaatctgc cttcaaatcc atgtttttac ccattgcatg tgattatgga
     1021 acctgggacc gaggagcagg aggagaacat tctaaattct gctcccatct tgtctttaca
     1081 tctcaggtca cttttagcaa agacagaccc ggacacttgc cattaatact acaggcttcc
     1141 ttcctcctac ccccttcccc caatcttatt catctcacct ctccagtagg tcgtggactc
     1201 atgcatttta taattaaaga aggccaaggc tcaggggtta agtgacttac tggaggttat
     1261 cagagaggaa gttgccaaac ccaggctagg aactgaatgg ttggtttaaa tccacccctc
     1321 tggcaggaga ctggggaata cacatgagcc gtgcagacag cagagggcag tcctgggggt
     1381 gggggcgcca gagggtttcc ggtacttttc agggcaattg aagttccggt cactactccc
     1441 ccccagagca ataagccaca tccggcgacg tgtggcaccc caccctggct gctacagatg
     1501 gggctggatg cagaagagaa ctccagctgg tccttaggga cacggagatc tgggtatata
     1561 atggaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc caaaaacata
     1621 aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga gcaactgcat
     1681 aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga tgcacatatc
     1741 gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc agaagctatg
     1801 aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa
     1861 ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc cgcgaacgac
     1921 atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac cgtggtgttc
     1981 gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc aatcatccaa
     2041 aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat gtacacgttc
     2101 gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga gtccttcgat
     2161 agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct gcctaaaggt
     2221 gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga tcctattttt
     2281 ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca tcacggtttt
     2341 ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt aatgtataga
     2401 tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag tgcgctgctg
     2461 gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata cgatttatct
     2521 aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg ggaagcggtt
     2581 gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga gactacatca
     2641 gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa agttgttcca
     2701 ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt taatcaaaga
     2761 ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa tccggaagcg
     2821 accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc ttactgggac
     2881 gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta caaaggctat
     2941 caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat cttcgacgca
     3001 ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg
     3061 gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca
     3121 accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa aggtcttacc
     3181 ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa gggcggaaag
     3241 atcgccgtgt aattctagag tcggggcggc cggccgcttc gagcagacat gataagatac
     3301 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3361 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3421 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc
     3481 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc cgtcgacccc gccaaccaga
     3541 tgatctgttc tcccgccaac cagatgatct gttctcgatg cccttgagag ccttcaaccc
     3601 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     3661 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg
     3721 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     3781 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3841 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4321 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     4801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4921 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5521 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5581 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgcgccct
     5641 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5701 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5761 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5821 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5881 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5941 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6001 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6061 ttaacaaaat attaacgttt acaatttccc attcgccatt caggctgcgc aactgttggg
     6121 aagggcgatc ggtgcgggcc tcttcgctat tacgccagcc caagctacca tgataagtaa
     6181 gtaatattaa ggtacgggag gtacttggag cggccgcaat aaaatatctt tattttcatt
     6241 acatctgtgt gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa
     6301 caaaacgaaa caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag
     6361 aacatttctc tatcgata
//
