LOCUS       sequence-14294-e        5468 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 5468)
  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 5468)
  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: pCSCre
FEATURES             Location/Qualifiers
     source          1..5468
                     /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             115..1146
                     /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"
     primer_bind     1490..1509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1627..1675
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(1701..1717)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1701..1717)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1764..1784)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1764..1782)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1803..1819)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1803..1819)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1816..1838)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1827..1843
                     /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(1851..1857,1858..1875,1876..1881))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1896..1917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2034..2051)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2205..2793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2285..2304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2964..3755,3756..3824))
                     /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     3587..3606
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3825..3929)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3955..4410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4092..4113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4304..4323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4484..5468
                     /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 atcccatcga ttcccgggaa ttcctgcagc cgatatgtcc
      121 aatttactga ccgtacacca aaatttgcct gcattaccgg tcgatgcaac gagtgatgag
      181 gttcgcaaga acctgatgga catgttcagg gatcgccagg cgttttctga gcatacctgg
      241 aaaatgcttc tgtccgtttg ccggtcgtgg gcggcatggt gcaagttgaa taaccggaaa
      301 tggtttcccg cagaacctga agatgttcgc gattatcttc tatatcttca ggcgcgcggt
      361 ctggcagtaa aaactatcca gcaacatttg ggccagctaa acatgcttca tcgtcggtcc
      421 gggctgccac gaccaagtga cagcaatgct gtttcactgg ttatgcggcg gatccgaaaa
      481 gaaaacgttg atgccggtga acgtgcaaaa caggctctag cgttcgaacg cactgatttc
      541 gaccaggttc gttcactcat ggaaaatagc gatcgctgcc aggatatacg taatctggca
      601 tttctgggga ttgcttataa caccctgtta cgtatagccg aaattgccag gatcagggtt
      661 aaagatatct cacgtactga cggtgggaga atgttaatcc atattggcag aacgaaaacg
      721 ctggttagca ccgcaggtgt agagaaggca cttagcctgg gggtaactaa actggtcgag
      781 cgatggattt ccgtctctgg tgtagctgat gatccgaata actacctgtt ttgccgggtc
      841 agaaaaaatg gtgttgccgc gccatctgcc accagccagc tatcaactcg cgccctggaa
      901 gggatttttg aagcaactca tcgattgatt tacggcgcta aggatgactc tggtcagaga
      961 tacctggcct ggtctggaca cagtgcccgt gtcggagccg cgcgagatat ggcccgcgct
     1021 ggagtttcaa taccggagat catgcaagct ggtggctgga ccaatgtaaa tattgtcatg
     1081 aactatatcc gtaacctgga tagtgaaaca ggggcaatgg tgcgcctgct ggaagatggc
     1141 gattagccat taacgcgtaa atgattgcta taattatttg atatttatgg tgacatatga
     1201 gaaaggattt caacatcgac ggaaaatatg tagtgctgtc tgtaagcact aatattcagt
     1261 cgccagccgt cattgtcact gtaaagctga gcgatagaat gcctgatatt gactcaatat
     1321 ccgttgcgtt tcctgtcaaa agtatgcgta gtgctgaaca tttcgtgatg aatgccaccg
     1381 aggaagaagc acggcgcggt tttgctaaag tgatgtctga gtttggcgaa ctcttgggta
     1441 aggttggaat tgtcgaggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     1501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     1561 atcgccgctc ccgattcgca gcgcatcgcc ttcnatcgcc ttcttgacga gttcttctga
     1621 ggggatcggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcggatc
     1681 cgtcgacctg cagcccgggg gatccactag ttctagagcg gccgcggcgc caatgcattg
     1741 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     1801 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     1861 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     1921 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     1981 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2041 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2101 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2161 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2221 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2281 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2341 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2401 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2461 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2521 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2581 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2641 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2701 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2761 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2821 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2881 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2941 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3001 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3061 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3121 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3181 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3241 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3301 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3361 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3421 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3481 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3541 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3601 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3661 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3721 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3781 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3841 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3901 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     3961 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4021 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     4081 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4141 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     4201 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4261 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4321 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4381 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     4441 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccata gccaattcaa
     4501 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa
     4561 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa
     4621 tatggcggac ttggcaccat gccaattcaa tatggcggac ttggcactgt gccaactggg
     4681 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc
     4741 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca
     4801 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac
     4861 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata
     4921 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc
     4981 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg
     5041 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc
     5101 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct
     5161 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc
     5221 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagt
     5281 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg
     5341 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc
     5401 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt
     5461 tagggaac
//
