LOCUS       sequence-431420-        3516 bp    DNA     circular SYN 15-APR-2025
DEFINITION  for stable fly cell lines; attL and attR sequences for genome 
            integration by phiC31, destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3516)
  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 3516)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRDB_051
FEATURES             Location/Qualifiers
     source          1..3516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     313..335
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     327..344
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     328..344
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    384..417
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     polyA_signal    886..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(897..916)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     951..970
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    1201..1234
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     primer_bind     complement(1295..1311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1295..1311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1308..1330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1319..1335
                     /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(1343..1349,1350..1367,1368..1373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1388..1409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1526..1543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1697..2285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1777..1796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2456..3247,3248..3316))
                     /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     3079..3098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3317..3421)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3489..3507
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
       61 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      121 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
      181 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      241 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      301 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
      361 cctcgcgaat gcatctagat cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
      421 ttcagaacgt cttcaccagt aatttacgtg ggaaaggtgc ttgtgtccca acaggctagg
      481 atataatgct gaaaccgtcc cccaagcgtt cagggtgggg tttgctacaa cttccgagtc
      541 caatgtgtcc gtgttcatga tatatatgct caagggcgag aattggacct agctttcgtg
      601 ttagtacgta gcatggtgac acaagcacag tagatcctgc ccgccgatcg tatatattaa
      661 gttaattctt atggaatata ataacatgtg gatggccagt ggtcggttgt tacacgccta
      721 ccgcgatgct gaatgacccg gactagagtg gcgaaattta tggcgtgtga cccgttatgc
      781 tccatttcgg tcagtgggtc attgctagta gtcgattgca ttgccattct ccgagtgatt
      841 tatggacccc agtgaagacg cattcggatc cgaattcaga catgataaga tacattgatg
      901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1081 acctctacaa atgtggtatg gctgattatg atcagtcgac cgatgccctt gagagccttc
     1141 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatggta
     1201 cgcgcccggg gagcccaagg gcacgccctg gcacccgatc ggatcccggg cccgtcgact
     1261 gcagaggcct gcatgcaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     1321 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     1381 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     1441 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1501 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1561 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1621 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1681 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1741 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1801 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1861 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1921 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1981 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2041 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2101 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2161 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2221 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2281 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2341 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2401 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2461 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2521 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2581 ctgcaatgat accgcgagat ccacgctcac cggctccaga tttatcagca ataaaccagc
     2641 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     2701 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     2761 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     2821 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     2881 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     2941 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3001 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3061 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3121 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3181 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3241 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3301 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     3361 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     3421 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     3481 cctataaaaa taggcgtatc acgaggccct ttcgtc
//
