LOCUS       sequence-211673-        3305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses eGFP donor in pUC19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3305)
  AUTHORS   Richardson CD, Kazane KR, Feng SJ, Zelin E, Bray NL, Schafer AJ, 
            Floor SN, Corn JE
  TITLE     CRISPR-Cas9 genome editing in human cells occurs via the Fanconi 
            anemia pathway.
  JOURNAL   Nat Genet. 2018 Aug;50(8):1132-1139. doi: 10.1038/s41588-018-0174-0.
            Epub 2018 Jul 27.
  PUBMED    30054595
REFERENCE   2  (bases 1 to 3305)
  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"; doi: 
            "10.1038/s41588-018-0174-0"; journalName: "Nat Genet"; date: 
            "2018-08"; volume: "50"; issue: "8"; pages: "1132-1139"
COMMENT     Sequence Label: pCR1075
FEATURES             Location/Qualifiers
     source          1..3305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(8..26)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        94..198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(199..267,268..1059)
                     /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"
     primer_bind     complement(417..436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1230..1818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1719..1738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1972..1989
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2106..2127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2142..2147,2148..2165,2166..2172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2180..2196
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     2185..2207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2204..2220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2204..2220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(2269..2271,2272..2274,2275..2787)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(2314..2335)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2575..2594)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(2909..2926)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2909..2925)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2918..2940)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3134..3153)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3253..3275
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
       61 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
      421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
      601 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
      661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
      721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
      781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
      841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1021 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     1081 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     1141 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     2041 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     2101 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     2161 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     2221 catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatccat ggtgagcaag
     2281 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2341 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     2401 ctgaagttca tctgcaccac cggcaagctg ccggtgccct ggcccaccct cgtgaccacc
     2461 ctgacgtacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2521 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     2581 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2641 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2701 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     2761 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     2821 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     2881 cagtccaagc tgagcaaaga ccgaattcac tggccgtcgt tttacaacgt cgtgactggg
     2941 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3001 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3061 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
     3121 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     3181 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     3241 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     3301 cgaga
//
