LOCUS       Exported                4151 bp ds-DNA     circular SYN 29-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4151)
  AUTHORS   Doudna Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 15, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4151
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          89..338
                     /organism="Escherichia virus T4"
                     /host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10665"
     source          371..620
                     /organism="Escherichia virus T4"
                     /host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10665"
     primer_bind     complement(39..89)
                     /label=oBA1728_BBR1ori_mob_F
     misc_feature    62..63
                     /label=BsaI OH
     misc_feature    64..84
                     /label=Pt4_common_region_4>5
     misc_feature    64..70
                     /label=AarI
     primer_bind     complement(70..89)
                     /label=oBA1651_pBFC0924_Gib_R
     misc_feature    75..78
                     /label=SMRT Gate OH Left
     misc_feature    79..80
                     /label=spacer in case extra trimming during AarI GG
     misc_feature    81..84
                     /label=pBBR1 Backbone Index (GACT)
     misc_feature    85..88
                     /label=4>5
     gene            89..338
                     /gene="rIIB"
                     /locus_tag="T4p278"
                     /label=rIIB
                     /db_xref="GeneID:1258618"
     CDS             89..338
                     /codon_start=1
                     /transl_table=11
                     /gene="rIIB"
                     /locus_tag="T4p278"
                     /product="rIIB protector from prophage-induced early
                     lysis"
                     /label=rIIB
                     /note="membrane integrity protector; mutants give rapid
                     lysis on various lysogenic strains due to effects of 
                     specific prophage products, mistakenly interpreted as 
                     related to lysis inhibition"
                     /db_xref="GeneID:1258618"
                     /protein_id="NP_049889.1"
                     /translation="EAKRPKVTISGDITVKVNSDAVIAPVAKSDIIWNASKKFISITVD
                     GVTYNATPNTHSNFQEILNLLVADKLEEAAQKINVRRA"
     misc_feature    89..338
                     /label=GT7_UP_Hom
     misc_feature    297..338
                     /label=Right_End_T4GT7_Deletion
     misc_feature    339..370
                     /label=BsmBI_HR_Dropout (UP is left, DN is right)
     misc_feature    339..342
                     /label=BsmBI_general_UP
     misc_feature    complement(344..349)
                     /label=BsmBI
     misc_feature    360..365
                     /label=BsmBI
     misc_feature    367..370
                     /label=BsmBI_DN
     misc_feature    371..620
                     /label=GT7_DN_Hom
     misc_feature    371..422
                     /label=Left_End_T4GT7_Deletion
     gene            371..418
                     /gene="52.1"
                     /locus_tag="T4p265"
                     /label=52.1
                     /note="erroneously identified as ac, has been identified
                     as hypothetical orf 52.1"
                     /db_xref="GeneID:1258635"
     CDS             371..418
                     /codon_start=1
                     /transl_table=11
                     /gene="52.1"
                     /locus_tag="T4p265"
                     /product="gp52.1 conserved hypothetical predicted membrane
                     protein"
                     /label=52.1
                     /db_xref="GeneID:1258635"
                     /protein_id="NP_049876.1"
                     /translation="LFISDVIMALHSLVW"
     gene            423..620
                     /gene="52"
                     /locus_tag="T4p264"
                     /label=52
                     /db_xref="GeneID:1258768"
     CDS             423..620
                     /codon_start=1
                     /transl_table=11
                     /gene="52"
                     /locus_tag="T4p264"
                     /product="DNA topisomerase II medium subunit"
                     /label=52
                     /note="gp52; bacteriophage T4 topoisomerase II is
                     comprised of gp39, gp52, and gp60 subunits; topisomerase II
                     catalyzes cleavage and re-ligation of double-stranded DNA"
                     /db_xref="GeneID:1258768"
                     /protein_id="NP_049875.1"
                     /translation="MQLNNRDLKSIIDNEALAYAMYTVENRAIPNMIDGFKPVQRFVIA
                     RALDLARGNKDKFHKLASIAG"
     primer_bind     621..669
                     /label=oBA1724_kanR_part4_gib_BBR1mob_R
     primer_bind     621..636
                     /label=oBA1650_pBFC0924_Gib_F
     misc_feature    621..624
                     /label=Pt4_common_region_6>4
     misc_feature    621..624
                     /label=6>4 OH
     misc_feature    join(625..4151,1..84)
                     /label=pBA707_p4
     misc_feature    625..639
                     /label=Pt4_common_region_6>4
     misc_feature    complement(625..628)
                     /label=SMRT Gate OH Right
     misc_feature    complement(633..639)
                     /label=AarI
     terminator      complement(640..666)
                     /label=KanR Terminator
     CDS             complement(667..1482)
                     /label=KanR
     primer_bind     complement(751..778)
                     /label=oBFC0064
     promoter        complement(1483..1630)
                     /label=KanR Promoter
     primer_bind     complement(1605..1664)
                     /label=oBA1723_kanR_part4_gib_BBR1mob_F
     misc_feature    1631..2931
                     /label=gbBA067
     misc_feature    complement(1631..1670)
                     /label=post-Mob from pFAB5526
     primer_bind     1631..1652
                     /label=oBA1725_gbBA067_BBR1mob_R
     CDS             complement(1671..2669)
                     /codon_start=1
                     /label=pFAB5526_MOB_recoded
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"
     misc_feature    complement(2670..2931)
                     /label=pre-MOB from pFAB5526
     misc_feature    2737..2788
                     /label=oriT
     misc_feature    2799..2819
                     /label=ACTUAL Sequence
     primer_bind     complement(2898..2931)
                     /label=oBA1726_gbBA067_BBR1mob_F
     primer_bind     2902..2945
                     /label=oBA1727_BBR1ori_mob_R
     misc_feature    join(2932..4151,1..63)
                     /label=from pBFC0924
     misc_feature    join(2932..4151,1..63)
                     /label=From pBBR1 backbone
     rep_origin      join(2932..4151,1..63)
                     /direction=RIGHT
                     /label=BBR1 ori
     CDS             join(3506..4151,1..17)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVLAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     complement(3664..3692)
                     /label=oBFC1872
     primer_bind     complement(3673..3692)
                     /label=oBFC1871
     primer_bind     3686..3710
                     /label=oBFC1870
     misc_feature    3688..3691
                     /label=BsaI OH
     misc_feature    3688
                     /label=silent c>t to remove BbsI
ORIGIN
        1 cgctgccgcg ccggtagcac ttgggttgcg cagcaacccg taagtgcgct gttccagact
       61 atccacctgc ccgaaccaga gacttatcag aagctaaacg ccctaaagtt actattagcg
      121 gtgatattac agttaaagtt aatagcgatg cagttattgc tccagttgct aaatctgaca
      181 ttatttggaa tgcatctaaa aaattcattt caattactgt tgatggcgta acttataacg
      241 caactcctaa tactcattca aacttccagg aaattcttaa tctgcttgta gcggataagt
      301 tggaagaagc ggcacaaaaa attaatgttc gtcgtgctga taagagacgg ctcaacgccc
      361 gtctcacagc cttttcattt ctgatgttat tatggcacta cattcactag tatggtaaat
      421 ttatgcaact gaataatcgc gatttaaaaa gtatcattga taatgaagca ttggcttatg
      481 ctatgtacac ggttgaaaat cgtgctatcc caaatatgat tgatggattt aagccagttc
      541 aacgatttgt tattgctcga gctcttgatt tggcacgagg aaataaagat aagtttcaca
      601 aactcgcttc tattgcaggt aggaggactt tggcaggtgg tgttacaacc aattaaccaa
      661 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
      721 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
      781 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
      841 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
      901 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
      961 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     1021 tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     1081 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     1141 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     1201 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     1261 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     1321 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     1381 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     1441 tcgcggcctg gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     1501 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     1561 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     1621 gaaggatcag cttgcgtttt cccttgtcca gatagcccag tagctgacat ttagcctaaa
     1681 tgacatttct ccggaaagtg cgcacggcca cgctcttgtt gacgttgagc ttgcttttga
     1741 cgttgaacct cgcgttgctc ggttaataat ttctcaccaa ccgctttaat aatggccgcc
     1801 gctttcgctt gcatgtcacg gttcagagga ccgagtgcat ctaataccgg tttcaaacgt
     1861 tcgcgtaaat cgcgcgcagt ttcctgtgcc tggtctgctt tcttacgcat ttcgcgggcg
     1921 ccggcagccg cttgcagtgc tggttcgtaa ccttgacgga ctgccttagt taagcgatcc
     1981 gcgactgcct caggggtttc cacacgttta gaaatgccta atttctccgc cagaccttgt
     2041 ggagcgtaag cacggggttc cacggcctga ggcgaaatcg taacatgtcc tacaggcggg
     2101 cgttcaagcg cttcataaaa tgcttggata cgagtatggc gcgctttact cccttcaatc
     2161 ccacgctgaa ggcccaggtc cgcaaccgca gcagcgaagg tcgtttgatc acgtgtcatt
     2221 tgggccttat taccaataaa ttctttagca ctcaagcgac catcttgggt caaggggact
     2281 acaaaggccg tcatatgtgg ggatgtctca tcgcgatgaa tcgaagctgt tacaatacgg
     2341 tcagcgccat atttatcggc taaccattta tgggctttct caaagaatgc tgcttgttgc
     2401 tcctgtgacg cgcttttcca ccactcgggc gaagcggtca tcacatattc tacggcaagt
     2461 actgcatctt tacggcgctt ttcaggcaac agttcacgca aacgacccat ggcctcgtcc
     2521 gtactactcg cggcccaatg ttcattttcc ggagtacgag atgcatccgc attcggtgtt
     2581 tcacgttcac gatacgcatg tttcagcgac gcagcaacat tacccatctt ggcgagtttc
     2641 ttacaacgca taattgcata agcggccatg cctgcccctc ccttttggtg tccaaccggc
     2701 tcgacggggg cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca
     2761 ctagactttg cttcgcaaag tcgtgaccgc ctacggcggc tgcgcctgcc cacgagcttg
     2821 accacaggga ttgcccaccg gctacccagc cttcgaccac atacccaccg gctccaactg
     2881 cgcggcctgc ggccttgccc catcaatttt tttaattttc tctggggaaa agcctccggc
     2941 ctgcggcctg cgcgcttcgc ttgccggttg gacaccaagt ggaaggcggg tcaaggctcg
     3001 cgcagcgacc gcgcagcggc ttggccttga cgcgcctgga acgacccaag cctatgcgag
     3061 tgggggcagt cgaagggcga agcccgcccg cctgcccccc gagcctcacg gcggcgagtg
     3121 cgggggttcc aagggggcag cgccaccttg ggcaaggccg aaggccgcgc agtcgatcaa
     3181 caagccccgg aggggccact ttttgccgga gggggagccg cgccgaaggc gtgggggaac
     3241 cccgcagggg tgcccttctt tgggcaccaa agaactagat atagggcgaa atgcgaaaga
     3301 cttaaaaatc aacaacttaa aaaagggggg tacgcaacag ctcattgcgg caccccccgc
     3361 aatagctcat tgcgtaggtt aaagaaaatc tgtaattgac tgccactttt acgcaacgca
     3421 taattgttgt cgcgctgccg aaaagttgca gctgattgcg catggtgccg caaccgtgcg
     3481 gcacccctac cgcatggaga taagcatggc cacgcagtcc agagaaatcg gcattcaagc
     3541 caagaacaag cccggtcact gggtgcaaac ggaacgcaaa gcgcatgagg cgtgggccgg
     3601 gcttattgcg aggaaaccca cggcggcaat gctgctgcat cacctcgtgg cgcagatggg
     3661 ccaccagaac gccgtggtgg tcagccaaaa gacactttcc aagctcatcg gacgttcttt
     3721 gcggacggtc caatacgcag tcaaggactt ggtggccgag cgctggatct ccgtcgtgaa
     3781 gctcaacggc cccggcaccg tgttggccta cgtggtcaat gaccgcgtgg cgtggggcca
     3841 gccccgcgac cagttgcgcc tgtcggtgtt cagtgccgcc gtggtggttg atcacgacga
     3901 ccaggacgaa tcgctgttgg ggcatggcga cctgcgccgc atcccgaccc tgtatccggg
     3961 cgagcagcaa ctaccgaccg gccccggcga ggagccgccc agccagcccg gcattccggg
     4021 catggaacca gacctgccag ccttgaccga aacggaggaa tgggaacggc gcgggcagca
     4081 gcgcctgccg atgcccgatg agccgtgttt tctggacgat ggcgagccgt tggagccgcc
     4141 gacacgggtc a
//
