LOCUS       sequence-11439-e        3893 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 3893)
  AUTHORS   Singla V, Hunkapiller J, Santos N, Seol AD, Norman AR, Wakenight P, 
            Skarnes WC, Reiter JF
  TITLE     Floxin, a resource for genetically engineering mouse ESCs.
  JOURNAL   Nat Methods. 2010 Jan . 7(1):50-2.
  PUBMED    19966808
REFERENCE   2  (bases 1 to 3893)
  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: "Nat 
            Methods. 2010 Jan . 7(1):50-2."
COMMENT     Sequence Label: pFloxin
FEATURES             Location/Qualifiers
     source          1..3893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      520..529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(532..565)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1045..1179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1082..1101)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1136..1155
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    1197..1244
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(1268..1284)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1268..1284)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1281..1303)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1292..1308
                     /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(1316..1322,1323..1340,1341..1346))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1361..1382
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1499..1516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1750..1769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2429..3220,3221..3289))
                     /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     3052..3071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3290..3394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3462..3480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3518..3540)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3640..3659
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3853..3875
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3867..3884
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3868..3884
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttcctt ctgcaggagc gtacagaacc cagggccctg gcacccgtgc agaccctggc
       61 ccaccccacc tgggcgctca gtgcccaaga gatgtccaca cctaggatgt cccgcggtgg
      121 gtggggggcc cgagagacgg gcaggccggg ggcaggcctg gccatgcggg gccgaaccgg
      181 gcactgccca gcgtggggcg cgggggccac ggcgcgcgcc cccagccccc gggcccagca
      241 ccccaaggcg gccaacgcca aaactctccc tcctcctctt cctcaatctc gctctcgctc
      301 tttttttttt tcgcaaaagg aggggagagg gggtaaaaaa atgctgcact gtgcggcgaa
      361 gccggtgagt gagcggcgcg gggccaatca gcgtgcgccg ttccgaaagt tgccttttat
      421 ggctcgagcg gccgcggcgg cgccctataa aacccagcgg cgcgacgcgc caccaccgcc
      481 gagaccgcgt ccgccccgcg agcacagagc ctcgcctttg ccaccatggc tataacttcg
      541 tatagcatac attatacgaa cggtagtgat aggcctttta gctacatctg ccaatccatc
      601 tcattttcac acacacacac accactttcc ttctggtcag tgggcacatg tccagcctca
      661 agtttatatc accaccccca atgcccaaca cttgtatggc cttgggcggg tcatcccccc
      721 ccccaccccc agtatctgca acctcaagct agcttgggtg cgttggttgt ggataagtag
      781 ctagactcca gcaaccagta acctctgccc tttctcctcc atgacaacca ggtaccggtg
      841 catgcggcgc gccactagtg atatcgctag cggatccacc acctacagcc cagtggggtc
      901 gctctagagc gacctcgagg gggggctaga taagtaatga tcataatcag ccatatcaca
      961 tctgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     1021 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     1081 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1141 ttgtccaaac tcatcaatgt atcttatcat gtctagatcc gggggtaccg cgtcgagaag
     1201 ttcctattcc gaagttccta ttctctagaa agtataggaa cttcgtcgac gcgaattcgt
     1261 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1321 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1381 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1441 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2101 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3421 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3481 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     3541 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     3601 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     3661 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3721 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3781 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3841 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctt
//
