LOCUS Exported 4639 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pNANOG-Luc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4639) AUTHORS Xu RH, Sampsell-Barron TL, Gu F, Root S, Peck RM, Pan G, Yu J, Antosiewicz-Bourget J, Tian S, Stewart R, Thomson JA TITLE NANOG is a direct target of TGFbeta/activin-mediated SMAD signaling in human ESCs. JOURNAL Cell Stem Cell. 2008 Aug 7. 3(2):196-206. PUBMED 18682241 REFERENCE 2 (bases 1 to 4639) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4639 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(63..80) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(234..822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(314..333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1022..1882) /codon_start=1 /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" polyA_signal 1987..2035 /note="synthetic polyadenylation signal" misc_feature 2049..2140 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 2089..2108 /label=RVprimer3 /note="pGL3 vector, forward primer" regulatory 2631..2640 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2637..4289 /codon_start=1 /product="firefly luciferase" /label=luciferase /note="synthetic luc2 version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" polyA_signal 4333..4454 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4344..4363) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4398..4417 /label=SV40pA-R /note="SV40 polyA, reverse primer" ORIGIN 1 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctcttccgct 61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 961 taaagtatat atgagtaaac ttggtctgac agcggccgca aatgctaaac cactgcagtg 1021 gttaccagtg cttgatcagt gaggcaccga tctcagcgat ctgcctattt cgttcgtcca 1081 tagtggcctg actccccgtc gtgtagatca ctacgattcg tgagggctta ccatcaggcc 1141 ccagcgcagc aatgatgccg cgagagccgc gttcaccggc ccccgatttg tcagcaatga 1201 accagccagc agggagggcc gagcgaagaa gtggtcctgc tactttgtcc gcctccatcc 1261 agtctatgag ctgctgtcgt gatgctagag taagaagttc gccagtgagt agtttccgaa 1321 gagttgtggc cattgctact ggcatcgtgg tatcacgctc gtcgttcggt atggcttcgt 1381 tcaactctgg ttcccagcgg tcaagccggg tcacatgatc acccatatta tgaagaaatg 1441 cagtcagctc cttagggcct ccgatcgttg tcagaagtaa gttggccgcg gtgttgtcgc 1501 tcatggtaat ggcagcacta cacaattctc ttaccgtcat gccatccgta agatgctttt 1561 ccgtgaccgg cgagtactca accaagtcgt tttgtgagta gtgtatacgg cgaccaagct 1621 gctcttgccc ggcgtctata cgggacaaca ccgcgccaca tagcagtact ttgaaagtgc 1681 tcatcatcgg gaatcgttct tcggggcgga aagactcaag gatcttgccg ctattgagat 1741 ccagttcgat atagcccact cttgcaccca gttgatcttc agcatctttt actttcacca 1801 gcgtttcggg gtgtgcaaaa acaggcaagc aaaatgccgc aaagaaggga atgagtgcga 1861 cacgaaaatg ttggatgctc atactcgtcc tttttcaata ttattgaagc atttatcagg 1921 gttactagta cgtctctcaa ggataagtaa gtaatattaa ggtacgggag gtattggaca 1981 ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt gtgtgaatcg 2041 atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag 2101 gctgtcccca gtgcaagtgc aggtgccaga acatttctct ggcctaactg gccggtacct 2161 gagctcgcta gcctcgaggc tggtttcaaa ctcctgactt caggtgatcc gcctgccacg 2221 gcctcccaat ttactgggat tacaggggtg ggccaccgcg cccggccttt ttcttaattt 2281 ttaaaaatat taaagtttta tcccattcct gttgaaccat attcctgatt taaaagttgg 2341 aaacgtggtg aacctagaag tatttgttgc tgggtttgtc ttcaggttct gttgctcggt 2401 tttctagttc cccacctagt ctgggttact ctgcagctac ttttgcatta caatggcctt 2461 ggtgagactg gtagacggga ttaactgaga attcacaagg gtgggtcagt agggggtgtg 2521 cccgccagga ggggtgggtc taaggtgata gagccttcat tataaatcta gagactccag 2581 gaagatctgg cctcggcggc caagcttggc aatccggtac tgttggtaaa gccaccatgg 2641 aagatgccaa aaacattaag aagggcccag cgccattcta cccactcgaa gacgggaccg 2701 ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc accatcgcct 2761 ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag atgagcgttc 2821 ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc gtggtgtgca 2881 gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc ggtgtggctg 2941 tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg ggcatcagcc 3001 agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac gtgcaaaaga 3061 agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac cagggcttcc 3121 aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag tacgacttcg 3181 tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt agtggcagta 3241 ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga ttcagtcatg 3301 cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc agcgtggtgc 3361 catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc ggctttcggg 3421 tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa gactataaga 3481 ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc actctcatcg 3541 acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg ctcagcaagg 3601 aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag ggctacggcc 3661 tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag cctggcgcag 3721 taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc ggtaagacac 3781 tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg agcggctacg 3841 ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg cacagcggcg 3901 acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg aagagcctga 3961 tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg ctgcaacacc 4021 ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc gagctgcccg 4081 ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc gtggactatg 4141 tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc gtggacgagg 4201 tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt ctcattaagg 4261 ccaagaaggg cggcaagatc gccgtgtaat aattctagag tcggggcggc cggccgcttc 4321 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa 4381 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct 4441 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg 4501 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc 4561 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc 4621 atgactatcg tcgccgcac //