LOCUS Exported 8781 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pShuttle-Cre-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8781) AUTHORS Jallepalli PV, Waizenegger IC, Bunz F, Langer S, Speicher MR, Peters JM, Kinzler KW, Vogelstein B, Lengauer C TITLE Securin is required for chromosomal stability in human cells. JOURNAL Cell. 2001 May 18. 105(4):445-57. PUBMED 11371342 REFERENCE 2 (bases 1 to 8781) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8781 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(11..28) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(182..770) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(262..281) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1801) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1564..1583 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1802..1906) /gene="bla" /label=AmpR promoter primer_bind 1974..1992 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" misc_signal 2223..2373 /label=Ad5 Psi /note="packaging signal for adenovirus serotype 5" enhancer 2672..3051 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3052..3255 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3206..3226 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 3252..3276 /label=LNCX /note="Human CMV promoter, forward primer" CDS 3297..4325 /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" polyA_signal 4405..4539 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4429..4448) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4483..4502 /label=SV40pA-R /note="SV40 polyA, reverse primer" CDS 8344..8535 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" misc_feature 8637..8777 /label=bom /note="basis of mobility region from pBR322" primer_bind 8692..8711 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 601 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta 661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag 1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac 1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc 1981 gtatcacgag gccctttcgt cttcaagaat tgatccgggc ccccatttcc cctcccttcc 2041 agctctctgc cccttttgga ttgaagccaa tatgataatg agggggtgga gtttgtgacg 2101 tggcgcgggg cgtgggaacg gggcgggtga cgtagtagtg tggcggaagt gtgatgttgc 2161 aagtgtggcg gaacacatgt aagcgacgga tgtggcaaaa gtgacgtttt tggtgtgcgc 2221 cggtgtacac aggaagtgac aattttcgcg cggttttagg cggatgttgt agtaaatttg 2281 ggcgtaaccg agtaagattt ggccattttc gcgggaaaac tgaataagag gaagtgaaat 2341 ctgaataatt ttgtgttact catagcgcgt aatctctagc atcgatgtcg agggatccct 2401 cgagtctaga gatatcgaat tcaagcttgt cgaccaattc tcatgtttga cagcttatca 2461 tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag 2521 atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa 2581 atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat 2641 attggctcat gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag 2701 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt 2761 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg 2821 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat 2881 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct 2941 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg 3001 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg 3061 ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc 3121 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa 3181 tgtcgtaata accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc 3241 tatataagca gagctcgttt agtgaaccgt cagatctcta gaagctgggt accaccatgt 3301 ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg 3361 aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct 3421 ggaaaatgct tctgtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga 3481 aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg 3541 gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt 3601 ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa 3661 aggaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt 3721 tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg 3781 catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg 3841 ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa 3901 cgctggttag caccgcaggt gtagagaagg cacttagcct gggggtaact aaactggtcg 3961 agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg 4021 tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg 4081 aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga 4141 gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg 4201 ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca 4261 tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg 4321 gcgattacga tgttccagat tacgcttacg atgttccaga ttacgctagc taaggatagc 4381 ggccgctcga ggccggcaag gccggatcca gacatgataa gatacattga tgagtttgga 4441 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt 4501 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat 4561 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac 4621 aaatgtggta tggctgatta tgatccggct gcctcgcgcg tttcggtgat gacggtgaaa 4681 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 4741 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga 4801 ggtcgactcg aagatctggg cgtggttaag ggtgggaaag aatatataag gtgggggtct 4861 tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc atgagcacca actcgtttga 4921 tggaagcatt gtgagctcat atttgacaac gcgcatgccc ccatgggccg gggtgcgtca 4981 gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg cccgcaaact ctactacctt 5041 gacctacgag accgtgtctg gaacgccgtt ggagactgca gcctccgccg ccgcttcagc 5101 cgctgcagcc accgcccgcg ggattgtgac tgactttgct ttcctgagcc cgcttgcaag 5161 cagtgcagct tcccgttcat ccgcccgcga tgacaagttg acggctcttt tggcacaatt 5221 ggattctttg acccgggaac ttaatgtcgt ttctcagcag ctgttggatc tgcgccagca 5281 ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt taaaacataa ataaaaaacc 5341 agactctgtt tggatttgga tcaagcaagt gtcttgctgt ctttatttag gggttttgcg 5401 cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg gtcctgtgta ttttttccag 5461 gacgtggtaa aggtgactct ggatgttcag atacatgggc ataagcccgt ctctggggtg 5521 gaggtagcac cactgcagag cttcatgctg cggggtggtg ttgtagatga tccagtcgta 5581 gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga ttgccagggg 5641 caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca tacgtgggga 5701 tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca tatccctccg 5761 gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg gaaatttgtc 5821 atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac ctccaagatt 5881 ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct gggcgaagat 5941 atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat aggccatttt 6001 tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg gcccaggggc 6061 gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg ggatcatgtc 6121 tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct gggaagaaag 6181 caggttcctg agcagctgcg acttaccgca gccggtgggc ccgtaaatca cacctattac 6241 cggctgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca ggggggccac 6301 ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca gaaggcgctc 6361 gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt tgagaccgtc 6421 cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc acagctcggt 6481 cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt ggggcggctt 6541 tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc tttccacggg 6601 cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc gggctgcgcg 6661 ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg gtcttcgccc 6721 tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgg 6781 cccttggcgc gcagcttgcc cttggaggag gcgccgcacg aggggcagtg cagacttttg 6841 agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc cgcgccgcag 6901 gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc ggggtcaaaa 6961 accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat gagccggtgt 7021 ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag aggcctgtcc 7081 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat 7141 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc 7201 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg 7261 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc 7321 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct 7381 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct 7441 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga 7501 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc taacttcgat 7561 cattggaccg ctgatcgtca cggcgattta tgccgcctcg gcgagcacat ggaacgggtt 7621 ggcatggatt gtaggcgccg ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc 7681 atggagccgg gccacctcga cctgaatgga agccggcggc acctcgctaa cggattcacc 7741 actccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca aaccaaccct 7801 tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg catctcgggc 7861 agcgttgggt cctggccacg ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta 7921 ggctggcggg gttgccttac tggttagcag aatgaatcac cgatacgcga gcgaacgtga 7981 agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa catgaatggt cttcggtttc 8041 cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct gcaccattat gttccggatc 8101 tgcatcgcag gatgctgctg gctaccctgt ggaacaccta catctgtatt aacgaagcgc 8161 tggcattgac cctgagtgat ttttctctgg tcccgccgca tccataccgc cagttgttta 8221 ccctcacaac gttccagtaa ccgggcatgt tcatcatcag taacccgtat cgtgagcatc 8281 ctctctcgtt tcatcggtat cattaccccc atgaacagaa atccccctta cacggaggca 8341 tcagtgacca aacaggaaaa aaccgccctt aacatggccc gctttatcag aagccagaca 8401 ttaacgcttc tggagaaact caacgagctg gacgcggatg aacaggcaga catctgtgaa 8461 tcgcttcacg accacgctga tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac 8521 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat 8581 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca 8641 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag 8701 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga 8761 gaaaataccg catcaggcgc t //