LOCUS       sequence-394486-        4703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mini circle middle entry plasmid for CreER-pA with flanking BsaI cut
            sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  AUTHORS   Kang Lab
  TITLE     mini-Golden version1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4703)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMC-ME_CreER-pA
FEATURES             Location/Qualifiers
     source          1..4703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..101)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     150..172
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     164..181
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     165..181
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             214..1242
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             1255..2190
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1303..2196
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    2226..2360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2250..2269)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2304..2323
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             2797..3588
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3681..4269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4170..4189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4423..4440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4599..4626)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
       61 cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcctggc agttccctac
      121 tctcgcgtta acgctagcat ggatgttttc ccagtcacga cgttgtaaaa cgacggccag
      181 tcttaagctc gggcccggtc tctagtcgcc accatgtcca atttactgac cgtacaccaa
      241 aatttgcctg cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac
      301 atgttcaggg atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc
      361 cggtcgtggg cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa
      421 gatgttcgcg attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag
      481 caacatttgg gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac
      541 agcaatgctg tttcactggt tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa
      601 cgtgcaaaac aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg
      661 gaaaatagcg atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac
      721 accctgttac gtatagccga aattgccagg atcagggtta aagatatctc acgtactgac
      781 ggtgggagaa tgttaatcca tattggcaga acgaaaacgc tggttagcac cgcaggtgta
      841 gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt
      901 gtagctgatg atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg
      961 ccatctgcca ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat
     1021 cgattgattt acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac
     1081 agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc
     1141 atgcaagctg gtggctggac caatgtaaat attgtcatga actatatccg taacctggat
     1201 agtgaaacag gggcaatggt gcgcctgctg gaagatggcg atctcgagcc atctgctgga
     1261 gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc taagaagaac
     1321 agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga tgctgagccc
     1381 cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc gatgatgggc
     1441 ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc gaagagggtg
     1501 ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg tgcctggcta
     1561 gagatcctga tgattggcct cgtctggcgc tccatggagc acccagtgaa gctactgttt
     1621 gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat ggtggagatc
     1681 ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca gggagaggag
     1741 tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt tctgtccagc
     1801 accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa gatcacagac
     1861 actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca ccagcggctg
     1921 gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg catggagcat
     1981 ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct ggaggcggcg
     2041 gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga ggagacggac
     2101 caaagccact tggccactgc gggctctact tcatcgcatt ccttgcaaaa gtattacatc
     2161 acgggggagg cagagggttt ccctgccaca gcttaatcta gaactatagt gagtcgtatt
     2221 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2281 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2341 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2401 gggaggtgtg ggaggttttt taattcctca agagacctct agaccagcca ggacagaaat
     2461 gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     2521 gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg
     2581 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     2641 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
     2701 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     2761 cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa
     2821 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     2881 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     2941 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     3001 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     3061 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     3121 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     3181 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     3241 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     3301 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     3361 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     3421 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     3481 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     3541 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac
     3601 ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     3661 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3721 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3781 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     3841 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3901 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     3961 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4021 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4081 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4141 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4201 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4261 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4321 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4381 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4441 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     4501 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4561 acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg
     4621 ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg
     4681 gccgttgctt cacaacgttc aaa
//
