LOCUS       sequence-41580-e        6120 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 6120)
  AUTHORS   Premsrirut PK, Dow LE, Kim SY, Camiolo M, Malone CD, Miething C, 
            Scuoppo C, Zuber J, Dickins RA, Kogan SC, Shroyer KR, Sordella R, 
            Hannon GJ, Lowe SW
  TITLE     A rapid and scalable system for studying gene function in mice using
            conditional RNA interference.
  JOURNAL   Cell. 2011 Apr 1;145(1):145-58.
  PUBMED    21458673
REFERENCE   2  (bases 1 to 6120)
  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."; 
            date: "2011-04-1"; volume: "145(1)"; pages: "145-58"
COMMENT     Sequence Label: pCol-TGM
FEATURES             Location/Qualifiers
     source          1..6120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(20..85,86..123)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    292..426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    546..680
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    join(692..710,711..733,734..752,753..775,776..794,795..817,
                     818..836,837..859,860..878,879..901,902..920,921..943,
                     944..962)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        995..1033
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1030..1054
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(1139..1141,1142..1144,1145..1858)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1139..1141,1142..1144,1145..1855)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1184..1205)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1445..1464)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1792..1813
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           1893..1986
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2000..2112
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     intron          2141..2713
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(3041..3629)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3121..3140)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3800..4591,4592..4660))
                     /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     4423..4442
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4661..4765)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4791..5246)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4928..4949)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5140..5159
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5373..5395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5387..5404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5388..5404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5414..5433
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5414..5432
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5465..5481
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5465..5481
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        5525..6024
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5560..5583)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5957..5976
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    6070..6117
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 cccccgggct gcagatctgt agggcgcagt agtccagggt ttccttgatg atgtcatact
       61 tatcctgtcc cttttttttc cacagctcgc ggttgaggac aaactcttcg cggtctttcc
      121 agtggggatc gacggtatcg ataagcttgc agatctgcga ctctagagga tctgcgactc
      181 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
      241 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
      301 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
      361 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      421 tggatctgcg actctagagg atcataatca gccataccac atttgtagag gttttacttg
      481 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
      541 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      601 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      661 tatcttatca tgtctggatc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      721 tcgagtttac cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct
      781 atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag
      841 tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
      901 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
      961 gaaaagtgaa agtcgagctc ggtacccggg tcgaggtagg cgtgtacggt gggaggccta
     1021 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt
     1081 tgacctccat agaagacacc gggaccgatc cagcctccgc ggccccgaat tgaccaccat
     1141 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1201 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1261 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1321 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1381 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1441 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     1501 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     1561 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     1621 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     1681 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     1741 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     1801 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg
     1861 aaccggtcga ctagggataa cagggtaatt gtttgaatga ggcttcagta ctttacagaa
     1921 tcgttgcctg cacatcttgg aaacacttgc tgggattact tcttcaggtt aacccaacag
     1981 aaggctcgag caaccagaat tcaaggggct actttaggag caattatctt gtttactaaa
     2041 actgaatacc ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa
     2101 ttaaatcact tttttcaatt cacgcgttga gaacttcagg gtgagtttgg ggacccttga
     2161 ttgttctttc tttttcgcta ttgtaaaatt catgttatat ggagggggca aagttttcag
     2221 ggtgttgttt agaatgggaa gatgtccctt gtatcaccat ggaccctcat gataattttg
     2281 tttctttcac tttctactct gttgacaacc attgtctcct cttattttct tttcattttc
     2341 tgtaactttt tcgttaaact ttagcttgca tttgtaacga atttttaaat tcacttttgt
     2401 ttatttgtca gattgtaagt actttctcta atcacttttt tttcaaggca atcagggtat
     2461 attatattgt acttcagcac agttttagag aacaattgtt ataattaaat gataaggtag
     2521 aatatttctg catataaatt ctggctggcg tggaaatatt cttattggta gaaacaacta
     2581 caccctggtc atcatcctgc ctttctcttt atggttacaa tgatatacac tgtttgagat
     2641 gaggataaaa tactctgagt ccaaaccggg cccctctgct aaccatgttc atgccttctt
     2701 ctctttccta cagctcctgg gcaacgtgct ggttgttgtg ctgtctcatc attttggcaa
     2761 aggattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2821 cctggctcac aaataccact gagatcgttt tccctctgcc aaaaattatg gggacatcat
     2881 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2941 gtgttggaat ttctcgatcg ctagcatcct aggcgcgcca catgtgagca aaaggccagc
     3001 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3061 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3121 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3181 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3241 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3301 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3361 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3421 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3481 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3541 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3601 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     3661 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     3721 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3781 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     3841 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     3901 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     3961 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4021 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4081 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4141 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     4201 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     4261 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     4321 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     4381 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     4441 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     4501 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     4561 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     4621 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     4681 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4741 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     4801 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4861 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4921 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4981 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5041 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5101 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5161 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5221 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
     5281 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     5341 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     5401 cagtgagcgc gcgtaatacg actcactata gggcgaattg gagctccacc gcggtggcgg
     5461 ccgctctaga actagtggat cccccgggct gcaggaatta attcgtcgac ctcgaaattc
     5521 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     5581 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     5641 aggcgccaac cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta
     5701 gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta
     5761 gcacgtctca ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc
     5821 ctttggggca gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga
     5881 aggggtgggt ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg
     5941 tcctccggag gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc
     6001 tcttcctcat ctccgggcct ttcgacctgc atccatctag atctcgatcg agcagctgaa
     6061 gcttatggag aagttactat tccgaagttc ctattctcta gaaagtatag gaacttcgat
//
