LOCUS       Exported                4817 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pgk-ATG-frt
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4817)
  AUTHORS   Beard C, Hochedlinger K, Plath K, Wutz A, Jaenisch R
  TITLE     Efficient method to generate single-copy transgenic mice by 
            site-specific integration in embryonic stem cells.
  JOURNAL   Genesis. 2006 Jan . 44(1):23-8.
  PUBMED    16400644
REFERENCE   2  (bases 1 to 4817)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4817
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     623..642
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        623..641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     674..690
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     674..690
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        734..1233
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(769..792)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1166..1185
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    1275..1322
                     /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)."
     misc_feature    1355..1458
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    1627..1761
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1881..2015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2027..2045
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2069..2087
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2111..2129
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2153..2171
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2204..2242
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2239..2263
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    2366..2487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2629..2647)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2668..2684)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2668..2684)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2681..2703)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2692..2708
                     /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(2716..2746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2761..2782
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2899..2916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3070..3658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3150..3169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3829..4689)
                     /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     4452..4471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4690..4794)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg agctccaccg
      661 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattaa ttcgtcgacc
      721 tcgaaattct accgggtagg ggaggcgctt ttcccaaggc agtctggagc atgcgcttta
      781 gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac acattccaca
      841 tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc accttctact
      901 cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa
      961 atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga gcaatggaag
     1021 cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt ctgggctcag
     1081 aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg cggggcgggc
     1141 gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg
     1201 ctgttctcct cttcctcatc tccgggcctt tcgacctgca tccatctaga tctcgagcag
     1261 ctgaagctta tggagaagtt actattccga agttcctatt ctctagaaag tataggaact
     1321 tcgatccccc tcgatccccc gggctgcaga tctgtagggc gcagtagtcc agggtttcct
     1381 tgatgatgtc atacttatcc tgtccctttt ttttccacag ctcgcggttg aggacaaact
     1441 cttcgcggtc tttccagtgg ggatcgacgg tatcgataag cttgcagatc tgcgactcta
     1501 gaggatctgc gactctagag gatcataatc agccatacca catttgtaga ggttttactt
     1561 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     1621 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     1681 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     1741 gtatcttatc atgtctggat ctgcgactct agaggatcat aatcagccat accacatttg
     1801 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     1861 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     1921 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1981 ccaaactcat caatgtatct tatcatgtct ggatcgagtt taccactccc tatcagtgat
     2041 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
     2101 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc
     2161 agtgatagag aaaagtgaaa gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg
     2221 ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
     2281 acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgcg gccccgaatt
     2341 cgatggccgc ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact
     2401 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     2461 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     2521 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa
     2581 atcggatcca ctagatctcg agggggggcc cggtaccagc ttttgttccc tttagtgagg
     2641 gttaatttcg agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     2701 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     2761 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     2821 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2881 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2941 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3001 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3841 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3901 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3961 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4021 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4081 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4141 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4201 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4261 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4321 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4381 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4441 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4501 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4561 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4621 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4681 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4741 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4801 tccccgaaaa gtgccac
//
