LOCUS        pBait        6207 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bait plasmid for PROBER (empty backbone encoding replaceable ccdB 
            cassette).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6207)
  AUTHORS   Mondal S, Ramanathan M, Miao W, Meyers RM, Rao D, Lopez-Pajares V, 
            Siprashvili Z, Reynolds DL, Porter DF, Ferguson I, Neela P, Zhao Y, 
            Meservey LM, Guo M, Yang YY, Li L, Wang Y, Khavari PA
  TITLE     PROBER identifies proteins associated with programmable 
            sequence-specific DNA in living cells.
  JOURNAL   Nat Methods. 2022 Aug;19(8):959-968. doi: 
            10.1038/s41592-022-01552-w. Epub 2022 Aug 4.
  PUBMED    35927480
REFERENCE   2  (bases 1 to 6207)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            //gt12365.cn
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-022-01552-w"; journalName: "Nat Methods"; date: 
            "2022-08"; volume: "19"; issue: "8"; pages: "959-968"
COMMENT     Sequence Label: pBait
FEATURES             Location/Qualifiers
     source          1..6207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..239
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     186..207
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     primer_bind     complement(1045..1062)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1051..1162
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1181..1201)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1186..1515
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             1575..2174
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1575..1594)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2071..2091
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2177..2298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2214..2233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2268..2287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2445..2462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2555..3143)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2635..2654)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3317..4111)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4112..4216)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4517..4539
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4531..4548
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4532..4548
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4559..4578
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4559..4577
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4582..4593
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(4713..5018)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(4815..4834)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(5360..6019)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5934..5953
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(6073..6079,6080..6097,6098..6103))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
ORIGIN
        1 ccggagtata aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag
       61 tgaacacgtc gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca
      121 atctgcagta aagtgcaagt taaagtgaat caattaaaag taaccagcaa ccaagtaaat
      181 caactgcaac tactgaaatc tgccaagaag taattattga atacaagaag agaactctga
      241 atagggaatt gggaattcgt taacagatcc gatatccgcc accaagcttc tgtctagagc
      301 cgccaccatg actgccctga ccgaaggtgc taagctgttt gagaaggaga ttccgtacat
      361 caccgagctg gaaggggacg tcgaaggaat gaagttcatc atcaagggag aaggaaccgg
      421 ggacgctacg actggaacca ttaaggccaa gtatatctgt accactggag atctgccagt
      481 gccttgggcc acccttgtgt caaccctctc gtatggagtg cagtgttttg ctaagtaccc
      541 tagccacatt aaggacttct tcaaatccgc catgccggaa ggttataccc aagagcgcac
      601 catttctttt gagggagatg gagtgtacaa gacccgcgcg atggtcacct atgagagggg
      661 atctatctac aaccgggtga ctctgactgg agaaaacttt aagaaggacg ggcatattct
      721 tcggaagaat gtcgccttcc agtgccctcc cagcatcctt tacattctcc ccgacactgt
      781 gaacaacgga atccgcgtgg agttcaatca agcctacgac atcgaggggg tgacggagaa
      841 gctggtgacc aagtgtagcc agatgaatcg gccactggcc ggttcagcgg ctgtccacat
      901 tccgcgctac catcatatca cttatcacac taagctctcc aaagaccgcg atgagaggag
      961 agatcacatg tgcctggtgg aagtggtcaa ggccgtcgat ctcgatacct atcagtaaac
     1021 ctgctctcaa aactgtcttc tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1081 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1141 aataaaatga ggaaattgca tcgaagacag tcagccttgt ggaatgtgtg tcagttaggg
     1201 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     1261 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     1321 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     1381 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     1441 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggt
     1501 ataggctttt gcaaaaagct ctctggctaa ctagagaacc cactgcttac tggcttatcg
     1561 tttaagccgc caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     1621 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     1681 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     1741 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     1801 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     1861 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     1921 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     1981 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     2041 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     2101 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     2161 agcccggtgc ctgaccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2221 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2281 actcatcaat gtatcttatc atgtcggatg agcggtatca gctcactcaa aggcccctcg
     2341 agggccttcc tttcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2401 catggtcata gctgtttcct tgcgtattgg gcgctctccg cttcctcgct cactgactcg
     2461 ctgcgctcgg tcgttcgggt aaagcctggg gtgcctaatg agcaaaaggc cagcaaaagg
     2521 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2581 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2641 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2701 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2761 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2821 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2881 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2941 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3001 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3061 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3121 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3181 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3241 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3301 cttggtctga cagttattag aaaaattcat ccagcagacg ataaaacgca atacgctggc
     3361 tatccggtgc cgcaatgcca tacagcacca gaaaacgatc cgcccattcg ccgcccagtt
     3421 cttccgcaat atcacgggtg gccagcgcaa tatcctgata acgatccgcc acgcccagac
     3481 ggccgcaatc aataaagccg ctaaaacggc cattttccac cataatgttc ggcaggcacg
     3541 catcaccatg ggtcaccacc agatcttcgc catccggcat gctcgctttc agacgcgcaa
     3601 acagctctgc cggtgccagg ccctgatgtt cttcatccag atcatcctga tccaccaggc
     3661 ccgcttccat acgggtacgc gcacgttcaa tacgatgttt cgcctgatga tcaaacggac
     3721 aggtcgccgg gtccagggta tgcagacgac gcatggcatc cgccataatg ctcacttttt
     3781 ctgccggcgc cagatggcta gacagcagat cctgacccgg cacttcgccc agcagcagcc
     3841 aatcacggcc cgcttcggtc accacatcca gcaccgccgc acacggaaca ccggtggtgg
     3901 ccagccagct cagacgcgcc gcttcatcct gcagctcgtt cagcgcaccg ctcagatcgg
     3961 ttttcacaaa cagcaccgga cgaccctgcg cgctcagacg aaacaccgcc gcatcagagc
     4021 agccaatggt ctgctgcgcc caatcatagc caaacagacg ttccacccac gctgccgggc
     4081 tacccgcatg caggccatcc tgttcaatca tactcttcct ttttcaatat tattgaagca
     4141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat
     4261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtggccgcta
     4381 cagggcgctc ccattcgcca ttcaggctgc gcaactgttg ggaagggcgt ttcggtgcgg
     4441 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     4501 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgacgta
     4561 atacgactca ctatagggcg aattgaagga aggccgtcaa ggccgcatac actgcggggg
     4621 ctctggagac gcgtcggagg acagtactcc gagcggagga cagtactccg agcggaggac
     4681 agtactccga agagcggtac cgagcctgac atttatattc cccagaacat caggttaatg
     4741 gcgtttttga tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag
     4801 accggcacac tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc
     4861 accgggtaaa gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc
     4921 accatccgtc gcccgggcgt gtcaataata tcactctgta catccacaaa cagacgataa
     4981 cggctctctc ttttataggt gtaaacctta aactgcattt caccagcccc tgttctcgtc
     5041 agcaaaagag ccgttcattt caataaaccg ggcgacctca gccatccctt cctgattttc
     5101 cgctttccag cgttcggcac gcagacgacg ggcttcattc tgcatggttg tgcttaccag
     5161 accggagata ttgacatcat atatgccttg agcaactgat agctgtcgct gtcaactgtc
     5221 actgtaatac gctgcttcat agcatacctc tttttgacat acttcgggta tacatatcag
     5281 tatatattct tataccgcaa aaatcagcgc gcaaatacgc atactgttat ctggctttta
     5341 gtaagccgga tccagatctt tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     5401 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     5461 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     5521 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     5581 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     5641 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     5701 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     5761 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     5821 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     5881 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     5941 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     6001 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgac ggatcctaac
     6061 tcaaaatcca cacattatac gagccggaag cataaagtgt aaagcctggg ggtaccgctc
     6121 ttcacggagg acagtactcc gagcggagga cagtactccg agcggaggac agtactccga
     6181 gcctctagac ggagactcta gcgagcg
//