LOCUS       sequence-14293-e        6429 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 6429)
  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 6429)
  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(T)
FEATURES             Location/Qualifiers
     source          1..6429
                     /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
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKRMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1522..2415
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     RMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     polyA_signal    2478..2612
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2502..2521)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2556..2575
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2725..2745)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2725..2743)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2764..2780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2764..2780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2777..2799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2788..2804
                     /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(2812..2818,2819..2836,2837..2842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2857..2878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2995..3012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3166..3754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3246..3265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3925..4716,4717..4785))
                     /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     4548..4567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4786..4890)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4916..5371)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5053..5074)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5265..5284
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5445..6429
                     /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 ccatggaaca cccggggaag ctcctgtttg ctcctaactt gctcctggac
     1861 aggaatcaag gtaaatgtgt ggaaggcatg 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 caacaaaaga atggagcatc tgtacagcat gaagtgcaag
     2221 aacgtggtgc ccctctatga cctgctgctg gagatgctgg acgcccaccg cctacatgcg
     2281 cccactagcc gtggaggggc atccgtggag gagacggacc aaagccactt ggccactgcg
     2341 ggctctactt catcgcattc cttgcaaaag tattacatca cgggggaggc agagggtttc
     2401 cctgccacag tctgagagct ccctggcgaa ttcaaggcct ctcgagcctc tagaactata
     2461 gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt ggacaaacca
     2521 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     2581 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     2641 ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt
     2701 gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2761 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2821 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2881 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2941 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3001 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3061 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3121 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3181 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3241 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3301 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3361 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3421 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3481 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3541 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3601 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3661 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3721 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3781 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3841 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3901 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3961 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4021 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4081 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4141 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4201 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4261 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4321 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4381 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4441 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4501 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4561 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4621 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4681 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4741 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4801 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4861 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     4921 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4981 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5041 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5101 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5161 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5221 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5281 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5341 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt
     5401 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat agccaattca
     5461 atatggcgta tatggactca tgccaattca atatggtgga tctggacctg tgccaattca
     5521 atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc agccaattca
     5581 atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg tgccaactgg
     5641 ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct gtgccaagtc
     5701 cgccatattg aattggcatg gtgccaataa tggcggccat attggctata tgccaggatc
     5761 aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc aggttcaata
     5821 ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat tgacgtagat
     5881 agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata ccgcccatag
     5941 ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga cgtcatatgg
     6001 gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt aaatggcccg
     6061 cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca atgggatggc
     6121 tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg gtaaatggcc
     6181 cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta ctattggaag
     6241 tacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat ggcccgcgat
     6301 ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca aatgggcgtt
     6361 ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag caatgctcgt
     6421 ttagggaac
//
