LOCUS Exported 3060 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Constitutive superfolder GFP (sfGFP) expression, controlled under E.coli hybrid J23117-J23106 promoter. ACCESSION . VERSION . KEYWORDS DT040A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3060) AUTHORS Toparlak OD, Zasso J, Bridi S, Serra MD, Macchi P, Conti L, Baudet ML, Mansy SS TITLE Artificial cells drive neural differentiation. JOURNAL Sci Adv. 2020 Sep 18;6(38). pii: 6/38/eabb4920. doi: 10.1126/sciadv.abb4920. Print 2020 Sep. PUBMED 32948587 REFERENCE 2 (bases 1 to 3060) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3060 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 283..302 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(521..625) /gene="bla" /label=AmpR promoter primer_bind 693..711 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" terminator 715..758 /label=bacterial terminator /note="putative bacterial transcription terminator" misc_feature 761..780 /label=BioBrick prefix /note="BioBrick prefix for parts that start with ""ATG""" CDS 845..868 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 869..1582 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 1715..1762 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" misc_feature 1764..1784 /label=BioBrick suffix /note="universal suffix for all parts" primer_bind complement(1790..1807) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1961..2549) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2041..2060) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2720..520) /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" ORIGIN 1 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 61 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 121 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 181 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 241 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 301 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 361 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 421 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 481 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 541 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 601 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag 661 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca 721 gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag 781 atttacggct agctcagtcc tagggattgt gctagcgata ctagaagaag gagaataatc 841 tatggactac aaagacgatg atgacaagat gcgtaaaggc gaagagctgt tcactggtgt 901 cgtccctatt ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg 961 cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg 1021 taaactgccg gtaccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt 1081 tgctcgttat ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg 1141 ctatgtgcag gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga 1201 agtgaaattt gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa 1261 agaagacggc aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta 1321 catcaccgcc gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt 1381 ggaggatggc agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg 1441 tcctgttctg ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc 1501 gaacgagaaa cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca 1561 tggtatggat gaactgtaca aataataacg actcaggctg ctacgcctgt gtactggaaa 1621 acaaaaccaa aacccaaaaa acaaaaaact gagcccattg gtatcgtgga aggactcgat 1681 caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaactagca taaccccttg gggcctctaa 1741 acgggtcttg aggggttttt tggtactagt agcggccgct gcaggcttcc tcgctcactg 1801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 1861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 1921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc 1981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 2041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 2101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 2161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 2221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 2281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 2341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 2401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 2461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 2521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 2581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 2641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 2701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 2761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 2821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 2881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 2941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 3001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt //