LOCUS Exported 5017 bp ds-DNA circular SYN 10-AGO-2015 DEFINITION . ACCESSION . VERSION . KEYWORDS pT3-Neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5017) AUTHORS Marco TITLE Direct Submission JOURNAL Exported segunda-feira, 17 de ago de 2015 from SnapGene Viewer 2.2.2 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5017 /organism="recombinant plasmid" /mol_type="other DNA" misc_feature 6..359 /note="IR/DR" primer_bind complement(404..420) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" protein_bind 428..444 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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(452..482) /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 497..518 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(806..1394) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1565..2425) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(2426..2530) /gene="bla" /note="AmpR promoter" primer_bind 3004..3020 /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" misc_feature 3040..3393 /note="IR/DR" promoter 3518..3903 /note="U3 MSCV" CDS 3914..4708 /codon_start=1 /note="Neo" /translation="MAEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(4743..4761) /note="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" polyA_signal 4787..5011 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" ORIGIN 1 gatccccttg aaatacatcc acaggtacac ctccaattga ctcaaatgat gtcaattagt 61 ctatcagaag cttctaaagc catgacatca ttttctggaa ttttccaagc tgtttaaagg 121 cacagtcaac ttagtgtatg taaacttctg acccactgga attgtgatac agtgaattat 181 aagtgaaata atctgtctgt aaacaattgt tggaaaaatg acttgtgtca tgcacaaagt 241 agatgtccta actgacttgc caaaactatt gtttgttaac aagaaatttg tggagtagtt 301 gaaaaacgag ttttaatgac tccaacttaa gtgtatgtaa acttccgact tcaactgtat 361 atctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg 421 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta 481 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg 541 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga 601 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg 661 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag 721 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc 781 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca 841 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt 901 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc 961 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc 1021 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc 1081 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact 1141 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg 1201 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta 1261 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca 1321 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa 1381 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg 1441 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc 1501 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 1561 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 1621 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 1681 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 1741 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 1801 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 1861 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 1921 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 1981 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 2041 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 2101 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 2161 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 2221 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 2281 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 2341 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 2401 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 2461 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag 2521 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca 2581 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 2641 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 2701 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 2761 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg 2821 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc 2881 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga 2941 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac 3001 gttgtaaaac gacggccagt gaattcgagc tcggtaccct acagttgaag tcggaagttt 3061 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt 3121 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca 3181 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc 3241 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt 3301 ccagaaaatg atgtcatggc tttagaagct tctgatagac taattgacat catttgagtc 3361 aattggaggt gtacctgtgg atgtatttca agggaattct gtggaatgtg tgtcagttag 3421 ggtgtggaaa gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatatcgat 3481 actagtttaa ttaagatctc gagcacgaga ctagccgtcg agggaatgaa agaccccacc 3541 tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac 3601 tgagaataga gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa 3661 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca 3721 gatgcggtcc cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa 3781 ggacctgaaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct 3841 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggcgcg 3901 ccgatccacc ggtatggccg aacaggatgg actgcacgca ggctcccccg ccgcttgggt 3961 ggaaagactg tttggatacg attgggctca gcagactatt ggatgctctg acgcagccgt 4021 gttccggctg agtgctcagg ggcgccccgt gctgtttgtc aagacagacc tgtcaggagc 4081 actgaacgag ctgcaggatg aagcagcacg actgagctgg ctggcaacca caggagtgcc 4141 ttgtgctgca gtcctggacg tggtcactga ggcaggacga gattggctgc tgctgggaga 4201 agtgccagga caggacctgc tgagctccca cctggcacca gctgagaagg tctccatcat 4261 ggcagatgcc atgcggagac tgcatactct ggaccccgcc acctgccctt tcgatcacca 4321 ggctaaacat cgcattgaga gggctcgcac ccgaatggaa gcaggcctgg tggaccagga 4381 cgatctggat gaggaacacc aggggctggc tcctgcagag ctgtttgcac ggctgaaagc 4441 cagaatgcca gacggagaag atctggtggt cacccatggc gacgcttgcc tgcccaacat 4501 catggtggag aatggcagat tcagtgggtt tattgattgt ggacgactgg gcgtcgccga 4561 cagataccag gatatcgccc tggctacaag agacattgca gaggaactgg gcggggaatg 4621 ggccgatagg ttcctggtgc tgtatggcat cgctgcccct gattcacaga ggattgcctt 4681 ttacagactg ctggatgagt tcttttgagc ggccgctcga gcatgcatct agagggccct 4741 attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt 4801 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 4861 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 4921 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 4981 aggcatgctg gggatgcggt gggctctatg gcttctg //