LOCUS       sequence-373960-        4864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of deGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4864)
  AUTHORS   Wandera KG, Alkhnbashi OS, Bassett HVI, Mitrofanov A, Hauns S, Migur
            A, Backofen R, Beisel CL
  TITLE     Anti-CRISPR prediction using deep learning reveals an inhibitor of 
            Cas13b nucleases.
  JOURNAL   Mol Cell. 2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003.
  PUBMED    35649413
REFERENCE   2  (bases 1 to 4864)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell. 
            2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003."
COMMENT     Sequence Label: P70a-deGFP-sc101-kan
FEATURES             Location/Qualifiers
     source          1..4864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      84..306
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(798..892)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(923..1717)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1034..1053)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1644..1663
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1896..1914
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     RBS             2252..2274
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2311..2332)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2572..2591)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2919..2940
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3225..3245
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3326..3412
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3950..4864,1..36))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt
       61 ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg
      121 agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag
      181 accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat
      241 aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta
      301 taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc
      361 tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg
      421 caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc
      481 gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca
      541 ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg
      601 gcttctcagg gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc
      661 agcagttcct gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca
      721 ttcagactgg ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta
      781 ctgtccctag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc gagcgttctg
      841 aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag tccaagcgag
      901 ctctcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
      961 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     1021 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     1081 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     1141 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     1201 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     1261 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     1321 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     1381 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     1441 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     1501 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     1561 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     1621 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     1681 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     1741 cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     1801 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccgacgtct
     1861 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     1921 gtcttcaaga attctggcga atcctctgac cagccagaaa acgacctttc tgtggtgaaa
     1981 ccggatgctg caattcagag cgccagcaag tgggggacag cagaagacct gaccgccgca
     2041 gagtggatgt ttgacatggt gaagactatc gcaccatcag ccagaaaacc gaattttgct
     2101 gggtgggcta acgatatccg cctgatgcgt gaacgtgacg gacgtaacca ccgcgacatg
     2161 tgtgtgctgt tccgctgggc atgctgagct aacaccgtgc gtgttgacaa ttttacctct
     2221 ggcggtgata atggttgcag ctagcaataa ttttgtttaa ctttaagaag gagatatacc
     2281 atggagcttt tcactggcgt tgttcccatc ctggtcgagc tggacggcga cgtaaacggc
     2341 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2401 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2461 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     2521 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2581 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2641 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2701 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     2761 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     2821 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2881 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2941 accgccgccg ggatctaact cgagcaaagc ccgccgaaag gcgggctttt ctgtgtcgac
     3001 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     3061 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     3121 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     3181 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     3241 gtaacagctg ctgggattac acatggcatg gatgaactat acaaataaat gtccagacct
     3301 gcaggcatgc aagctctaga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     3361 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgcc
     3421 ctagacctag ggtacgggtt ttgctgcccg caaacgggct gttctggtgt tgctagtttg
     3481 ttatcagaat cgcagatccg gcttcagccg gtttgccggc tgaaagcgct atttcttcca
     3541 gaattgccat gattttttcc ccacgggagg cgtcactggc tcccgtgttg tcggcagctt
     3601 tgattcgata agcagcatcg cctgtttcag gctgtctatg tgtgactgtt gagctgtaac
     3661 aagttgtctc aggtgttcaa tttcatgttc tagttgcttt gttttactgg tttcacctgt
     3721 tctattaggt gttacatgct gttcatctgt tacattgtcg atctgttcat ggtgaacagc
     3781 tttgaatgca ccaaaaactc gtaaaagctc tgatgtatct atctttttta caccgttttc
     3841 atctgtgcat atggacagtt ttccctttga tatgtaacgg tgaacagttg ttctactttt
     3901 gtttgttagt cttgatgctt cactgataga tacaagagcc ataagaacct cagatccttc
     3961 cgtatttagc cagtatgttc tctagtgtgg ttcgttgttt ttgcgtgagc catgagaacg
     4021 aaccattgag atcatactta ctttgcatgt cactcaaaaa ttttgcctca aaactggtga
     4081 gctgaatttt tgcagttaaa gcatcgtgta gtgtttttct tagtccgtta tgtaggtagg
     4141 aatctgatgt aatggttgtt ggtattttgt caccattcat ttttatctgg ttgttctcaa
     4201 gttcggttac gagatccatt tgtctatcta gttcaacttg gaaaatcaac gtatcagtcg
     4261 ggcggcctcg cttatcaacc accaatttca tattgctgta agtgtttaaa tctttactta
     4321 ttggtttcaa aacccattgg ttaagccttt taaactcatg gtagttattt tcaagcatta
     4381 acatgaactt aaattcatca aggctaatct ctatatttgc cttgtgagtt ttcttttgtg
     4441 ttagttcttt taataaccac tcataaatcc tcatagagta tttgttttca aaagacttaa
     4501 catgttccag attatatttt atgaattttt ttaactggaa aagataaggc aatatctctt
     4561 cactaaaaac taattctaat ttttcgcttg agaacttggc atagtttgtc cactggaaaa
     4621 tctcaaagcc tttaaccaaa ggattcctga tttccacagt tctcgtcatc agctctctgg
     4681 ttgctttagc taatacacca taagcatttt ccctactgat gttcatcatc tgagcgtatt
     4741 ggttataagt gaacgatacc gtccgttctt tccttgtagg gttttcaatc gtggggttga
     4801 gtagtgccac acagcataaa attagcttgg tttcatgctc cgttaagtca tagcgactaa
     4861 tcgc
//
