LOCUS       sequence-267595-        7357 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 7357)
  AUTHORS   Lin CC, Liu LZ, Addison JB, Wonderlin WF, Ivanov AV, Ruppert JM
  TITLE     A KLF4-miRNA-206 autoregulatory feedback loop can promote or inhibit
            protein translation depending upon cell context.
  JOURNAL   Mol Cell Biol. 2011 Jun;31(12):2513-27. Epub 2011 Apr 25.
  PUBMED    21518959
REFERENCE   2  (bases 1 to 7357)
  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: "Mol Cell 
            Biol."; date: "2011-06"; volume: "31(12)"; pages: "2513-27. Epub 
            2011 Apr 25"
COMMENT     Sequence Label: pBpuro - HA KLF4 FL
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1377..1403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             1377..1403
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3680..3700)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3685..4014
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3865..4000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4024..4623
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4024..4043)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4520..4540
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4844..5313
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5443..5460)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5614..6202)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5694..5713)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6362..7153,7154..7222))
                     /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     6985..7004
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7223..7327)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc cggcgagtct gacatgtacc
     1381 cctacgacgt gcccgactac gccgctgtca gcgacgcgct gctcccatct ttctccacgt
     1441 tcgcgtctgg cccggcggga agagagaaga cactgcgtca agcaggtgcc ccgaataacc
     1501 gctggcggga ggagctctcc cacatgaagc gacttccccc agtgcttccc ggccgcccct
     1561 atgacctggc ggcggcgacc gtggccacag acctggagag cggcggagcc ggtgcggctt
     1621 gcggcggtag caacctggcg cccctacctc ggagagagac cgaggagttc aacgatctcc
     1681 tggacctgga ctttattctc tccaattcgc tgacccatcc tccggagtca gtggccgcca
     1741 ccgtgtcctc gtcagcgtca gcctcctctt cgtcgtcgcc gtcgagcagc ggccctgcca
     1801 gcgcgccctc cacctgcagc ttcacctatc cgatccgggc cgggaacgac ccgggcgtgg
     1861 cgccgggcgg cacgggcgga ggcctcctct atggcaggga gtccgctccc cctccgacgg
     1921 ctcccttcaa cctggcggac atcaacgacg tgagcccctc gggcggcttc gtggccgagc
     1981 tcctgcggcc agaattggac ccggtgtaca ttccgccgca gcagccgcag ccgccaggtg
     2041 gcgggctgat gggcaagttc gtgctgaagg cgtcgctgag cgcccctggc agcgagtacg
     2101 gcagcccgtc ggtcatcagc gtcagcaaag gcagccctga cggcagccac ccggtggtgg
     2161 tggcgcccta caacggcggg ccgccgcgca cgtgccccaa gatcaagcag gaggcggtct
     2221 cttcgtgcac ccacttgggc gctggacccc ctctcagcaa tggccaccgg ccggctgcac
     2281 acgacttccc cctggggcgg cagctcccca gcaggactac cccgaccctg ggtcttgagg
     2341 aagtgctgag cagcagggac tgtcaccctg ccctgccgct tcctcccggc ttccatcccc
     2401 acccggggcc caattaccca tccttcctgc ccgatcagat gcagccgcaa gtcccgccgc
     2461 tccattacca agagctcatg ccacccggtt cctgcatgcc agaggagccc aagccaaaga
     2521 ggggaagacg atcgtggccc cggaaaagga ccgccaccca cacttgtgat tacgcgggct
     2581 gcggcaaaac ctacacaaag agttcccatc tcaaggcaca cctgcgaacc cacacaggtg
     2641 agaaacctta ccactgtgac tgggacggct gtggatggaa attcgcccgc tcagatgaac
     2701 tgaccaggca ctaccgtaaa cacacggggc accgcccgtt ccagtgccaa aaatgcgacc
     2761 gagcattttc caggtcggac cacctcgcct tacacatgaa gaggcatttt taaatcccag
     2821 acagtggata tgacccacac tgccagaaga gaattcagta ttttttactt ttcacactgt
     2881 cttcccgatg agggaaggag cccagccaga aagcactaca atcatggtca agttcccaac
     2941 tgagtcatct tgtgagtgga taatcaggaa aaatgaggaa tccaaaagac aaaaatcaaa
     3001 gaacagatgg ggtctgtgac tggatcttct atcattccaa ttctaaatcc gacttgaata
     3061 ttcctggact tacaaaatgc caagggggtg actggaagtt gtggatatca gggtataaat
     3121 tatatccgtg agttggggga gggaagacca gaattccctt gaattgtgta ttgatgcaat
     3181 ataagcataa aagatcacct tgtattctct ttaccttcta aaagccatta ttatgatgtt
     3241 agaagaagag gaagaaattc aggtacagaa aacatgttta aatagcctaa atgatggtgc
     3301 ttggtgagtc ttggttctaa aggtaccaaa caaggaagcc aaagttttca aactgctgca
     3361 tactttgaca aggaaaatct atatttgtct tccgatcaac atttatgacc taagtcaggt
     3421 aatatacctg gtttacttct ttagcatttt tatgcagaca gtctgttatg cactgtggtt
     3481 tcagatgtgc aataatttgt acaatggttt attcccaagt atgccttaag cagaacaaat
     3541 gtgtttttct atatagttcc ttgccttaat aaatatgtaa tataaattta aaaaaaaaaa
     3601 aaaaaaaaac tcgacctgag taactgacac aatggatcgg gtcgagtcta gaattcgcca
     3661 gcacagtggt cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3721 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3781 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3841 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3901 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3961 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     4021 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4081 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4141 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4201 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4261 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4321 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4381 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4441 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4501 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4561 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     4621 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     4681 ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt
     4741 aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg
     4801 agaatagaga agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac
     4861 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct
     4921 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     4981 acagatggtc cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt
     5041 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
     5101 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
     5161 ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata
     5221 aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg
     5281 agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg
     5341 gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg
     5401 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5461 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5521 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5581 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5641 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5701 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5761 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5821 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5881 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5941 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6001 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6061 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6121 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6181 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6241 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6301 ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6361 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6421 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6481 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6541 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6601 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6661 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6721 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6781 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6841 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6901 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6961 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7021 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7081 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7141 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7201 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7261 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7321 ttccgcgcac atttccccga aaagtgccac ctgcagc
//
