LOCUS       Exported                4530 bp ds-DNA     circular SYN 09-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4530)
  AUTHORS   Brady Cress
  TITLE     Direct Submission
  JOURNAL   Exported Monday, May 9, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4530
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1448..2748
                     /organism="gbBA067_BBR1_mob_recoded"
     primer_bind     complement(103..125)
                     /label=oBFC0451
     primer_bind     239..260
                     /label=oBFC1567
     primer_bind     complement(244..265)
                     /label=oBFC0453
     primer_bind     complement(268..291)
                     /label=oBFC1617
     primer_bind     complement(278..300)
                     /label=oBFC1710
     terminator      301..429
                     /label=BBa_B0015 Terminator
     primer_bind     301..332
                     /label=oBFC0174
     primer_bind     301..324
                     /label=oBFC1711
     primer_bind     309..330
                     /label=oBFC0177
     primer_bind     complement(337..380)
                     /label=oBFC0177
     primer_bind     complement(361..387)
                     /label=oBFC1189
     primer_bind     complement(403..429)
                     /label=oBFC0696
     primer_bind     complement(406..430)
                     /label=oBFC0055
     primer_bind     complement(406..430)
                     /label=oBFC0806
     primer_bind     complement(406..429)
                     /label=oBFC0422
     primer_bind     complement(406..429)
                     /label=oBFC0426
     primer_bind     complement(406..429)
                     /label=oBFC0488
     primer_bind     complement(406..429)
                     /label=oBFC0490
     primer_bind     complement(407..450)
                     /label=oBA1653_RFP924_droupout_Gib_R
     primer_bind     complement(408..456)
                     /label=oBA1708_RFPdropout_part4_gib_R
     primer_bind     complement(408..444)
                     /label=oBFC1287
     primer_bind     complement(408..430)
                     /label=oBFC0309
     primer_bind     complement(408..430)
                     /label=oBFC1335
     primer_bind     complement(408..430)
                     /label=oBFC0785
     primer_bind     complement(408..430)
                     /label=oBFC0356
     primer_bind     complement(408..429)
                     /label=oBFC0692
     primer_bind     complement(408..423)
                     /label=oBFC1196
     primer_bind     410..453
                     /label=oBA1650_pBFC0924_Gib_F
     primer_bind     complement(412..430)
                     /label=oBFC1204
     primer_bind     complement(412..430)
                     /label=oBFC0738
     primer_bind     complement(422..450)
                     /label=oBFC1476
     misc_feature    430..4064
                     /label=pBA707_p4
     primer_bind     430..486
                     /label=oBA1724_kanR_part4_gib_BBR1mob_R
     misc_feature    430..456
                     /label=Pt4_common_region_6>4
     primer_bind     complement(435..456)
                     /label=oBFC1462
     misc_feature    438..441
                     /label=6>4 OH
     misc_feature    complement(442..445)
                     /label=SMRT Gate OH Right
     misc_feature    complement(450..456)
                     /label=AarI
     terminator      complement(457..483)
                     /label=KanR Terminator
     CDS             complement(484..1299)
                     /label=KanR
     primer_bind     complement(568..595)
                     /label=oBFC0064
     promoter        complement(1300..1447)
                     /label=KanR Promoter
     primer_bind     complement(1422..1481)
                     /label=oBA1723_kanR_part4_gib_BBR1mob_F
     misc_feature    1448..2748
                     /label=gbBA067
     misc_feature    complement(1448..1487)
                     /label=post-Mob from pFAB5526
     primer_bind     1448..1469
                     /label=oBA1725_gbBA067_BBR1mob_R
     CDS             complement(1488..2486)
                     /codon_start=1
                     /label=pFAB5526_MOB_recoded
                     /translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
                     AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
                     DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
                     DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
                     VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
                     LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLG"
     misc_feature    complement(2487..2748)
                     /label=pre-MOB from pFAB5526
     misc_feature    2554..2605
                     /label=oriT
     misc_feature    2616..2636
                     /label=ACTUAL Sequence
     primer_bind     complement(2715..2748)
                     /label=oBA1726_gbBA067_BBR1mob_F
     primer_bind     2719..2762
                     /label=oBA1727_BBR1ori_mob_R
     rep_origin      2749..4031
                     /direction=RIGHT
                     /label=BBR1 ori
     misc_feature    2749..4031
                     /label=From pBBR1 backbone
     misc_feature    2749..4031
                     /label=from pBFC0924
     CDS             3323..3985
                     /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(3481..3509)
                     /label=oBFC1872
     primer_bind     complement(3490..3509)
                     /label=oBFC1871
     primer_bind     3503..3527
                     /label=oBFC1870
     misc_feature    3505..3508
                     /label=BsaI OH
     misc_feature    3505
                     /label=silent c>t to remove BbsI
     primer_bind     complement(4007..4064)
                     /label=oBA1728_BBR1ori_mob_F
     misc_feature    4030..4031
                     /label=BsaI OH
     primer_bind     4032..4091
                     /label=oBA1707_RFPdropout_BBR1_part4_gib_F
     misc_feature    4032..4064
                     /label=Pt4_common_region_4>5
     misc_feature    4032..4038
                     /label=AarI
     primer_bind     complement(4038..4084)
                     /label=oBA1651_pBFC0924_Gib_R
     misc_feature    4043..4046
                     /label=SMRT Gate OH Left
     primer_bind     4045..4095
                     /label=oBA1652_RFP924_droupout_Gib_F
     misc_feature    4047..4048
                     /label=spacer in case extra trimming during AarI GG
     misc_feature    4049..4052
                     /label=pBBR1 Backbone Index (GACT)
     primer_bind     4053..4092
                     /label=oBFC1286
     misc_feature    4053..4056
                     /label=4>5 OH
                     /label=4b(3)
     misc_feature    4053..4056
                     /label=BsaI OH
     primer_bind     4064..4092
                     /label=oBFC0308
     primer_bind     4064..4092
                     /label=oBFC0355
     primer_bind     4064..4092
                     /label=oBFC1361
     primer_bind     4064..4086
                     /label=oBFC1705
     promoter        4065..4118
                     /label=Bba_R0040 TetR Promoter
     primer_bind     4065..4092
                     /label=oBFC0784
     primer_bind     4119..4147
                     /label=oBFC1616
     RBS             4119..4143
                     /label=mRFP1 Ribsosome Binding Site
     misc_feature    4138
                     /label=pt mut makes higher RFP expression
     CDS             join(4144..4530,1..291)
                     /codon_start=1
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(4145..4164)
                     /label=oBFC0275
     primer_bind     4162..4184
                     /label=oBFC0189
     primer_bind     4162..4184
                     /label=oBFC0380
     primer_bind     4162..4184
                     /label=oBFC0391
     primer_bind     4176..4195
                     /label=oBFC0277
     primer_bind     complement(4177..4201)
                     /label=oBFC0523
     primer_bind     complement(4177..4199)
                     /label=oBFC0521
     primer_bind     complement(4177..4198)
                     /label=oBFC0439
     primer_bind     complement(4186..4217)
                     /label=oBFC0535
     primer_bind     complement(4186..4217)
                     /label=oBFC0187
     primer_bind     complement(4186..4216)
                     /label=oBFC0551
     primer_bind     complement(4186..4216)
                     /label=oBFC0559
     primer_bind     complement(4186..4216)
                     /label=oBFC0390
     primer_bind     complement(4186..4215)
                     /label=oBFC0543
     primer_bind     complement(4186..4215)
                     /label=oBFC0379
     primer_bind     4186..4214
                     /label=oBFC0186
     primer_bind     4187..4216
                     /label=oBFC0289
     primer_bind     4197..4224
                     /label=oBFC0191
     primer_bind     4197..4224
                     /label=oBFC0381
     primer_bind     4197..4224
                     /label=oBFC0392
     primer_bind     complement(4201..4222)
                     /label=oBFC0475
     primer_bind     4201..4219
                     /label=oBFC0528
     primer_bind     complement(4213..4235)
                     /label=oBFC1637
     primer_bind     complement(4220..4241)
                     /label=oBFC0441
     primer_bind     complement(4240..4269)
                     /label=oBFC0602
     primer_bind     complement(4249..4270)
                     /label=oBFC0443
     misc_feature    4266
                     /label=t-->c in Qi coli
     primer_bind     4267..4295
                     /label=oBFC0601
     primer_bind     4267..4289
                     /label=oBFC0197
     primer_bind     4267..4289
                     /label=oBFC0384
     primer_bind     4267..4289
                     /label=oBFC0395
     primer_bind     complement(4268..4287)
                     /label=oBFC0467
     primer_bind     complement(4270..4290)
                     /label=oBFC0291
     primer_bind     complement(4298..4319)
                     /label=oBFC0445
     primer_bind     4337..4360
                     /label=oBFC0301
     primer_bind     4363..4373
                     /label=oBFC1058_fixed
     primer_bind     4363..4373
                     /label=oBFC1060_fixed
     primer_bind     4363..4373
                     /label=oBFC1062_fixed
     primer_bind     4363..4373
                     /label=oBFC1447
     primer_bind     complement(4383..4404)
                     /label=oBFC0447
     primer_bind     complement(4417..4442)
                     /label=oBFC0472
     primer_bind     complement(4417..4442)
                     /label=oBFC0473
     primer_bind     complement(4417..4442)
                     /label=oBFC0474
     primer_bind     complement(4476..4497)
                     /label=oBFC0449
ORIGIN
        1 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
       61 atgtacccgg aggacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
      121 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
      181 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga ggactacacc
      241 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataaggatct
      301 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
      361 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
      421 gcgtttatag aagaccaagg aggactttgg caggtggtgt tacaaccaat taaccaattc
      481 tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      541 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      601 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      661 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      721 gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg
      781 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      841 ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
      901 cgaatgcaac cggcgcagga acactgcaag cgcatcaaca atattttcac ctgaatcagg
      961 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     1021 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     1081 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1141 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     1201 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     1261 cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     1321 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     1381 tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg ctgagttgaa
     1441 ggatcagctt gcgttttccc ttgtccagat agcccagtag ctgacattta gcctaaatga
     1501 catttctccg gaaagtgcgc acggccacgc tcttgttgac gttgagcttg cttttgacgt
     1561 tgaacctcgc gttgctcggt taataatttc tcaccaaccg ctttaataat ggccgccgct
     1621 ttcgcttgca tgtcacggtt cagaggaccg agtgcatcta ataccggttt caaacgttcg
     1681 cgtaaatcgc gcgcagtttc ctgtgcctgg tctgctttct tacgcatttc gcgggcgccg
     1741 gcagccgctt gcagtgctgg ttcgtaacct tgacggactg ccttagttaa gcgatccgcg
     1801 actgcctcag gggtttccac acgtttagaa atgcctaatt tctccgccag accttgtgga
     1861 gcgtaagcac ggggttccac ggcctgaggc gaaatcgtaa catgtcctac aggcgggcgt
     1921 tcaagcgctt cataaaatgc ttggatacga gtatggcgcg ctttactccc ttcaatccca
     1981 cgctgaaggc ccaggtccgc aaccgcagca gcgaaggtcg tttgatcacg tgtcatttgg
     2041 gccttattac caataaattc tttagcactc aagcgaccat cttgggtcaa ggggactaca
     2101 aaggccgtca tatgtgggga tgtctcatcg cgatgaatcg aagctgttac aatacggtca
     2161 gcgccatatt tatcggctaa ccatttatgg gctttctcaa agaatgctgc ttgttgctcc
     2221 tgtgacgcgc ttttccacca ctcgggcgaa gcggtcatca catattctac ggcaagtact
     2281 gcatctttac ggcgcttttc aggcaacagt tcacgcaaac gacccatggc ctcgtccgta
     2341 ctactcgcgg cccaatgttc attttccgga gtacgagatg catccgcatt cggtgtttca
     2401 cgttcacgat acgcatgttt cagcgacgca gcaacattac ccatcttggc gagtttctta
     2461 caacgcataa ttgcataagc ggccatgcct gcccctccct tttggtgtcc aaccggctcg
     2521 acgggggcag cgcaaggcgg tgcctccggc gggccactca atgcttgagt atactcacta
     2581 gactttgctt cgcaaagtcg tgaccgccta cggcggctgc gcctgcccac gagcttgacc
     2641 acagggattg cccaccggct acccagcctt cgaccacata cccaccggct ccaactgcgc
     2701 ggcctgcggc cttgccccat caattttttt aattttctct ggggaaaagc ctccggcctg
     2761 cggcctgcgc gcttcgcttg ccggttggac accaagtgga aggcgggtca aggctcgcgc
     2821 agcgaccgcg cagcggcttg gccttgacgc gcctggaacg acccaagcct atgcgagtgg
     2881 gggcagtcga agggcgaagc ccgcccgcct gccccccgag cctcacggcg gcgagtgcgg
     2941 gggttccaag ggggcagcgc caccttgggc aaggccgaag gccgcgcagt cgatcaacaa
     3001 gccccggagg ggccactttt tgccggaggg ggagccgcgc cgaaggcgtg ggggaacccc
     3061 gcaggggtgc ccttctttgg gcaccaaaga actagatata gggcgaaatg cgaaagactt
     3121 aaaaatcaac aacttaaaaa aggggggtac gcaacagctc attgcggcac cccccgcaat
     3181 agctcattgc gtaggttaaa gaaaatctgt aattgactgc cacttttacg caacgcataa
     3241 ttgttgtcgc gctgccgaaa agttgcagct gattgcgcat ggtgccgcaa ccgtgcggca
     3301 cccctaccgc atggagataa gcatggccac gcagtccaga gaaatcggca ttcaagccaa
     3361 gaacaagccc ggtcactggg tgcaaacgga acgcaaagcg catgaggcgt gggccgggct
     3421 tattgcgagg aaacccacgg cggcaatgct gctgcatcac ctcgtggcgc agatgggcca
     3481 ccagaacgcc gtggtggtca gccaaaagac actttccaag ctcatcggac gttctttgcg
     3541 gacggtccaa tacgcagtca aggacttggt ggccgagcgc tggatctccg tcgtgaagct
     3601 caacggcccc ggcaccgtgt tggcctacgt ggtcaatgac cgcgtggcgt ggggccagcc
     3661 ccgcgaccag ttgcgcctgt cggtgttcag tgccgccgtg gtggttgatc acgacgacca
     3721 ggacgaatcg ctgttggggc atggcgacct gcgccgcatc ccgaccctgt atccgggcga
     3781 gcagcaacta ccgaccggcc ccggcgagga gccgcccagc cagcccggca ttccgggcat
     3841 ggaaccagac ctgccagcct tgaccgaaac ggaggaatgg gaacggcgcg ggcagcagcg
     3901 cctgccgatg cccgatgagc cgtgttttct ggacgatggc gagccgttgg agccgccgac
     3961 acgggtcacg ctgccgcgcc ggtagcactt gggttgcgca gcaacccgta agtgcgctgt
     4021 tccagactat ccacctgccc gaaccagaga cttatcatgt cttctcccta tcagtgatag
     4081 agattgacat ccctatcagt gatagagata ctgagcacgg atctgaaaga ggagaaatga
     4141 tctatggcga gtagcgaaga tgttatcaaa gagttcatgc gtttcaaagt tcgtatggaa
     4201 ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg tccgtacgaa
     4261 ggtactcaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt cgcttgggac
     4321 atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc ggctgacatc
     4381 ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt tatgaacttc
     4441 gaggacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg tgagttcatc
     4501 tacaaagtta aactgcgtgg tactaacttc
//
