LOCUS Exported 3783 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Col1a-3'probe SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3783) 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 3783) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3783 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(892..908) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(893..909) /label=pBluescriptKS /note="For pBluescript vector" primer_bind complement(938..958) /label=T3 /note="T3 promoter, forward primer" promoter complement(938..956) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(977..993) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(977..993) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(990..1012) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1001..1017 /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(1025..1055) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1070..1091 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1208..1225) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1379..1967) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1459..1478) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2138..2998) /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 2761..2780 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2999..3103) /gene="bla" /label=AmpR promoter rep_origin complement(3129..3584) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3266..3287) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3478..3497 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3711..3733 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3725..3742 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3726..3742 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3752..3771 /label=T7 /note="T7 promoter, forward primer" promoter 3752..3770 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ccaccgcggt ggcggccgct ctagataaat tttcctttca cgttgtcctc atgcaaagtg 61 ctaataataa attattaaag gccatcatta atagtcttgg atactccgtg accatatgaa 121 cgtgtcctat gggtgccaca gtagactggt agcacaagag agggaagcta caaatggcct 181 gttccttcct ttcaaatttg agacctgtct ccccgagtga aaggggtcat ctgtgtcctt 241 atgtctactg tctggttcac cccagacttc tgcctgctgg attctcgggc cttctactat 301 gtcaccgtcc ccattctctg agatgtgggg gacaattagt taccatccct ctagggcctc 361 aggttctcag atggacagag ggagagaggg aaaaggaaga agaaaaagca aaaagttttg 421 gggataaagg aaaaggaaag aagggggcag gaagaagggg ggaacaattc cacgggctcc 481 ctgtaccgag ccagcatgct atcgtcccgg gcacctctgt gatgtggggt cctgtccttt 541 ctggtgacac cccggaagtt ccaccacagc tctctaagat cccccgggga gagttccttg 601 agggctgggc acctctctct ggtgagatca agggtgaccc aagaaattct catcctcctt 661 gtgacacccc agcattagaa ggccccttct atactaaatt atggtttctt tgggctagag 721 gtggggagag gccagctacc cctccatgtg tgaccaagga gcaacctggt cctccatgtc 781 tcagtccctg tttctgctgc ttgaatccct ttgagggctc atgaacctcc caggatccat 841 gggcaacacc tcccaggcct caggtctgca ggaattcgat atcaagctta tcgataccgt 901 cgacctcgag ggggggcccg gtacccagct tttgttccct ttagtgaggg ttaattgcgc 961 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 1021 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 1081 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 1141 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 1201 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 1261 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 1321 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 1381 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 1441 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 1501 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 1561 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 1621 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 1681 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 1741 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 1801 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct 1861 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 1921 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 1981 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 2041 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 2101 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 2161 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 2221 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 2281 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 2341 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 2401 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 2461 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 2521 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 2581 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 2641 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 2701 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 2761 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 2821 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 2881 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 2941 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 3001 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 3061 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 3121 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 3181 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 3241 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 3301 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 3361 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 3421 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 3481 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 3541 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag 3601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 3661 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 3721 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattgga 3781 gct //