LOCUS       sequence-431455-        3100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Selection) surface marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3100)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 3100)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA104
FEATURES             Location/Qualifiers
     source          1..3100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1010..1026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1010..1026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1034..1050
                     /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)."
     promoter        complement(join(1058..1064,1065..1082,1083..1088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1103..1124
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1241..1258)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1412..2000)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1492..1511)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2178..2987)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2901..2920
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2979..2998
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta ggagccacca tgaatcccgc catcagcgtg gctctgctgc
      481 tgagcgtgct gcaagtgtct aggggccaga aagtcacctc tctgactgct tgtctggtga
      541 accagaatct gaggctggat tgtaggcacg agaacaacac caaggacaac agcatccagc
      601 acgagttctc tctgactagg gaaaagagga agcacgtgct gagcggcact ctgggcattc
      661 ccgaacacac ctataggtct agggtcaccc tcagcaatca gccctacatc aaggtgctga
      721 ccctcgccaa cttcaccacc aaggatgagg gcgattactt ctgtgagctg agggtgagcg
      781 gcgccaaccc catgagcagc aacaagtcca tcagcgtgta tagggacaag ctggtgaagt
      841 gcggcggcat ctctctgctg gtgcagaata ctagctggat gctgctgctg ctgctgtctc
      901 tgtctctgct ccaagctctg gacttcatct ctctgtgagg aagcacgtct tcatcggatc
      961 ccgggcccgt cgactgcaga ggcctgcatg caagcttggt gtaatcatgg tcatagctgt
     1021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1141 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     1201 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     1261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     1321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     1381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     1441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     1501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     1561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     1621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     1681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     1741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     1801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     1861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     1921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     1981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     2041 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     2101 agatcctttt aaattaaaaa tgaagtttta aatcaagccc aatctgaata atgttacaac
     2161 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2221 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2281 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2341 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2401 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     2461 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2521 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     2581 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     2641 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     2701 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     2761 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     2821 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     2881 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     2941 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     3001 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     3061 atgtaacatc agagattttg agacacgggc cagagctgca
//
