LOCUS       sequence-14343-e        6392 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 6392)
  AUTHORS   Werdien D, Peiler G, Ryffel GU
  TITLE     FLP and Cre recombinase function in Xenopus embryos.
  JOURNAL   Nucleic Acids Res. 2001 Jun 1. 29(11):E53-3.
  PUBMED    11376165
REFERENCE   2  (bases 1 to 6392)
  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: "Nucleic 
            Acids Res. 2001 Jun 1. 29(11):E53-3."
COMMENT     Sequence Label: pCS_FLPER(T2)
FEATURES             Location/Qualifiers
     source          1..6392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             94..1362
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=FLP
                     /note="FLP is a site-specific recombinase from 
                     Saccharomyces cerevisiae. Recombination occurs at FRT 
                     sequences."
                     /translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIASCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     CDS             1474..2409
                     /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             1522..2412
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     polyA_signal    2441..2575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2465..2484)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2519..2538
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2688..2708)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2688..2706)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2727..2743)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2743)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2740..2762)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2751..2767
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2775..2781,2782..2799,2800..2805))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2820..2841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2958..2975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3129..3717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3209..3228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3888..4679,4680..4748))
                     /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     4511..4530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4749..4853)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4879..5334)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5016..5037)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5228..5247
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5408..6392
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccacagcc accatgccac aatttgatat attatgtaaa
      121 acaccaccta aggtcctggt tcgtcagttt gtggaaaggt ttgaaagacc ttcaggggaa
      181 aaaatagcat catgtgctgc tgaactaacc tatttatgtt ggatgattac tcataacgga
      241 acagcaatca agagagccac attcatgagc tataatacta tcataagcaa ttcgctgagt
      301 ttcgatattg tcaacaaatc actccagttt aaatacaaga cgcaaaaagc aacaattctg
      361 gaagcctcat taaagaaatt aattcctgct tgggaattta caattattcc ttacaatgga
      421 caaaaacatc aatctgatat cactgatatt gtaagtagtt tgcaattaca gttcgaatca
      481 tcggaagaag cagataaggg aaatagccac agtaaaaaaa tgcttaaagc acttctaagt
      541 gagggtgaaa gcatctggga gatcactgag aaaatactaa attcgtttga gtatacctcg
      601 agatttacaa aaacaaaaac tttataccaa ttcctcttcc tagctacttt catcaattgt
      661 ggaagattca gcgatattaa gaacgttgat ccgaaatcat ttaaattagt ccaaaataag
      721 tatctgggag taataatcca gtgtttagtg acagagacaa agacaagcgt tagtaggcac
      781 atatacttct ttagcgcaag gggtaggatc gatccacttg tatatttgga tgaatttttg
      841 aggaattctg aaccagtcct aaaacgagta aataggaccg gcaattcttc aagcaacaaa
      901 caggaatacc aattattaaa agataactta gtcagatcgt acaacaaggc tttgaagaaa
      961 aatgcgcctt atccaatctt tgctataaag aatggcccaa aatctcacat tggaagacat
     1021 ttgatgacct catttctgtc aatgaagggc ctaacggagt tgactaatgt tgtgggaaat
     1081 tggagcgata agcgtgcttc tgccgtggcc aggacaacgt atactcatca gataacagca
     1141 atacctgatc actacttcgc actagtttct cggtactatg catatgatcc aatatcaaag
     1201 gaaatgatag cattgaagga tgagactaat ccaattgagg agtggcagca tatagaacag
     1261 ctaaagggta gtgctgaagg aagcatacga taccccgcat ggaatgggat aatatcacag
     1321 gaggtactag actacctttc atcctacata aatagacgca tatccgtacg cggatccatg
     1381 aaaggtggga tacgaaaaga ccgaagagga gggagaatgt tgaaacacaa gcgccagaga
     1441 gatgatgggg agggcagggg tgaagtgggg tctgctggag acatgagagc tgccaacctt
     1501 tggccaagcc cgctcatgat caaacgctct aagaagaaca gcctggcctt gtccctgacg
     1561 gccgaccaga tggtcagtgc cttgttggat gctgagcccc ccatactcta ttccgagtat
     1621 gatcctacca gacccttcag tgaagcttcg atgatgggct tactgaccaa cctggcagac
     1681 agggagctgg ttcacatgat caactgggcg aagagggtgc caggctttgt ggatttgacc
     1741 ctccatgatc aggtccacct tctagaatgt gcctggctag agatcctgat gattggtctc
     1801 gtctggcgct ccatggagca cccagtgaag ctactgtttg ctcctaactt gctcttggac
     1861 aggaaccagg gaaaatgtgt agagggcatg gtggagatct tcgacatgct gctggctaca
     1921 tcatctcggt tccgcatgat gaatctgcag ggagaggagt ttgtgtgcct caaatctatt
     1981 attttgctta attctggagt gtacacattt ctgtccagca ccctgaagtc tctggaagag
     2041 aaggaccata tccaccgagt cctggacaag atcacagaca ctttgatcca cctgatggcc
     2101 aaggcaggcc tgaccctgca gcagcagcac cagcggctgg cccagctcct cctcatcctc
     2161 tcccacatca ggcacatgag taacaaaggc atggagcatc tgtacagcat gaagtgcaag
     2221 aacgtggtgc ccctctatga cctgctgctg gaggcggcgg acgcccaccg cctacatgcg
     2281 cccactagcc gtggaggggc atccgtggag gagacggacc aaagccactt ggccactgcg
     2341 ggctctactt catcgcattc cttgcaaaag tattacatca cgggggaggc agagggtttc
     2401 cctgccacag tgctagaact atagtgagtc gtattacgta gatccagaca tgataagata
     2461 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2521 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2581 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaatt
     2641 cgcggccgcg gcgccaatgc attgggcccg gtacccagct tttgttccct ttagtgaggg
     2701 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     2761 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     2821 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2881 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2941 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3541 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     3841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     3901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     3961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4741 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4801 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     4861 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4921 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4981 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     5041 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     5101 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     5161 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5221 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5281 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt
     5341 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5401 gccagtcgac catagccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt
     5461 ggatctggac ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt
     5521 ggatctggac cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc
     5581 ggacttggca ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag
     5641 gggtcttggc cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc
     5701 catattggct atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat
     5761 atggctattg gccaggttca atactatgta ttggccctat gccatatagt attccatata
     5821 tgggttttcc tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg
     5881 gggcttccta ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg
     5941 tctttcctat tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga
     6001 cgtcaattac ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc
     6061 accattgacg tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt
     6121 gacgtcaatg acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt
     6181 ggcaagtaca ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc
     6241 aatggcggta aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag
     6301 gggcaatgac gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg
     6361 aggtctatat aagcaatgct cgtttaggga ac
//
