LOCUS       sequence-13260-e        6545 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 6545)
  AUTHORS   Dejosez M, Levine SS, Frampton GM, Whyte WA, Stratton SA, Barton MC,
            Gunaratne PH, Young RA, Zwaka TP
  TITLE     Ronin/Hcf-1 binds to a hyperconserved enhancer element and regulates
            genes involved in the growth of embryonic stem cells.
  JOURNAL   Genes Dev. 2010 Jul 15. 24(14):1479-84.
  PUBMED    20581084
REFERENCE   2  (bases 1 to 6545)
  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: "Genes Dev. 
            2010 Jul 15. 24(14):1479-84."
COMMENT     Sequence Label: pEF1a-Ronin-FLAG-IRES-Neo
FEATURES             Location/Qualifiers
     source          1..6545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..1181
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          250..1172
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1129..1149
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1220..1352
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1335..1354
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1397..1415
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    2439..3011
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2462..3013
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2606..2623)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2833..2852
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3013..3810
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3070..3089)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3680..3699
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3874..3922
                     /note="synthetic polyadenylation signal"
     rep_origin      4017..4472
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4104..4123)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4314..4335
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4541..4560)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4660..4682
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4720..4738)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4806..4910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4911..4979,4980..5771)
                     /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(5129..5148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5942..6530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6431..6450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaattcaag cttcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
       61 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
      121 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
      181 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
      241 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
      301 ccttgcgtgc cttgaattac ttccacctgg ctccagtacg tgattcttga tcccgagctg
      361 gagccagggg cgggccttgc gctttaggag ccccttcgcc tcgtgcttga gttgaggcct
      421 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      481 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      541 ggcaagatag tcttgtaaat gcgggccagg atctgcacac tggtatttcg gtttttgggc
      601 ccgcggccgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      661 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      721 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      781 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tccagggggc tcaaaatgga
      841 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
      901 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
      961 attagttctg gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1021 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1081 aattctcctt ggaatttggc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1141 cagtggttca aagttttttt cttccatttc aggtgtcgtg aacacgggct gcagaagttg
     1201 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     1261 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     1321 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1381 ttaaggctag agtacttaat acgactcact ataggctagc ctcgagatct gccaccatgc
     1441 ctggctttac gtgctgcgtg ccaggctgct acaacaactc gcaccgggac aaggcgctgc
     1501 acttctacac gtttccaaag gacgctgagt tgcggcgcct ctggctcaag aacgtgtcgc
     1561 gtgccggcgt cagtgggtgc ttctccacct tccagcccac cacaggccac cgtctctgca
     1621 gcgttcactt ccagggcggc cgcaagacct acacggtacg cgtccccacc atcttcccgc
     1681 tgcgcggcgt caatgagcgc aaagtagcgc gcagacccgc tggggccgcg gccgcccgcc
     1741 gcaggcagca gcagcaacag cagcagcagc agcaacagca gcaacagcag cagcagcagc
     1801 aacagcagca gcagcagcag cagcagcagt cctcaccctc tgcctccact gcccagactg
     1861 cccagctgca gccgaacctg gtatctgctt ccgcggccgt gcttctcacc cttcaggcca
     1921 ctgtagacag cagtcaggct ccgggatccg tacagccggc gcccatcact cccactggag
     1981 aagacgtgaa gcccatcgat ctcacagtgc aagtggagtt tgcagccgca gagggcgcag
     2041 ccgctgcggc cgccgcgtcg gagttacagg ctgctaccgc agggctggag gctgccgagt
     2101 gccctatggg cccccagttg gtggtggtag gggaagaggg cttccctgat actggctccg
     2161 accattcgta ctccttgtcg tcaggcacca cggaggagga gctcctgcgc aagctgaatg
     2221 agcagcggga catcctggct ctgatggaag tgaagatgaa agagatgaaa ggcagcattc
     2281 gccacctgcg tctcactgag gccaagctgc gcgaagaact gcgtgagaag gatcggctgc
     2341 ttgccatggc tgtcatccgc aagaagcacg gaatggatta caaggatgac gataagtgat
     2401 ctagagtcga cccgggcggc cgctctagcc caattccgcc cctctccctc ccccccccct
     2461 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt
     2521 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg
     2581 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc
     2641 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt
     2701 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta
     2761 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg
     2821 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag
     2881 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag
     2941 tcgaggttaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     3001 cacgatgata atatgggcat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3061 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3121 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3181 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3241 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3301 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3361 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3421 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3481 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3541 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3601 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3661 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3721 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3781 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3841 aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt ttcattacat
     3901 ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tccgggctgg cgtaatagcg
     3961 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc
     4021 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4081 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4141 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagagc
     4201 tttacggcac ctcgaccgca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     4261 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     4321 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     4381 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa tatttaacgc
     4441 gaattttaac aaaatattaa cgtttacaat ttcgcctgat gcggtatttt ctccttacgc
     4501 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     4561 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4621 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4681 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4741 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4801 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5041 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5101 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5161 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5221 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5281 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5341 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5401 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5461 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5521 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5581 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5641 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5701 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5761 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5821 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5881 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5941 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6001 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6061 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6421 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6481 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6541 acgcg
//
