LOCUS       Exported                5847 bp ds-DNA     circular SYN 06-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5847)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Aug 19, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     SnapGene NotesSequence Type: SyntheticConfirmed Experimentally: 
            0Transformed Into: unspecifiedSnapGene UUID: 
            cecdf57d-c56c-4a53-9a61-b3625bc0b25dSequence Document Type: 
            UNAImported FromPath: 
            /Users/benjaminadler/Library/CloudStorage/GoogleDrive-ba14@berkeley.
            edu/My Drive/BA_Doudna_Lab/BA_DNA_DB/BA_SnapGene/DNA 
            Files/Phage_Immune_SystemsFilename: 
            pBA1751_pJEX__mCpolTA_ECOR31_STANDARD.dna
FEATURES             Location/Qualifiers
     source          1..5847
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      253..475
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     primer_bind     complement(935..957)
                     /label=oBFC0813a
     primer_bind     complement(935..957)
                     /label=oBFC0813
     primer_bind     complement(941..990)
                     /label=oBA1709_SC101_part4_gib_F
     misc_feature    957..959
                     /label=BsmBI_OH
     primer_bind     961..1020
                     /label=oBA1718_kanR_part4_gib_R
     misc_feature    961..990
                     /label=Part4_Ori_AbR_Common
     terminator      complement(991..1017)
                     /label=KanR Terminator
     primer_bind     991..1016
                     /label=oBFC0313
     CDS             complement(1018..1833)
                     /label=KanR
     primer_bind     complement(1102..1129)
                     /label=oBFC0064
     promoter        complement(1834..1981)
                     /label=KanR Promoter
     primer_bind     1982..2035
                     /label=oBA2268_pJEX-SC101Gib_F
     misc_feature    1982..2006
                     /label=Pt4_common_region_4>5
     primer_bind     complement(1982..2006)
                     /label=oBA2260_pt4_common_4>5_R
     misc_feature    1982..1988
                     /label=AarI
     primer_bind     complement(1988..2006)
                     /label=oBA1651_pBFC0924_Gib_R
     misc_feature    1993..1996
                     /label=SMRT Gate OH Left
     misc_feature    1997..1998
                     /label=spacer in case extra trimming during AarI GG
     misc_feature    1999..2002
                     /label=pBBR1 Backbone Index (GACT)
     misc_feature    2003..2006
                     /label=4>5 OH
                     /label=4b(3)
     misc_feature    2003..2006
                     /label=BsaI OH
     misc_feature    2007..2957
                     /label=from gbBA189
     misc_feature    2007..2010
                     /label=1>2
     CDS             complement(2017..2595)
                     /codon_start=1
                     /transl_table=0
                     /label=eilR
                     /translation="MGYLNREERRETIMQAAMRVALDQGFTGMTVRNIATAAGVAAGQV
                     HHHFTSSGELKSQAFIRVIREMMDLQRLSRTAGWREQLFSALGSEDGRLEPYIRLWRQA
                     QLLADSDPEIKSAYLLTMNLWHDEAVRIIRAGHAAGEFTLRDSAENIAWRLISLVCGLD
                     GIYVLGMPEVDDAAFTRHLQHVIQLELFS"
     misc_feature    2053
                     /label=AarI removal
     misc_feature    2596..2610
                     /label=GSG_RBS
     RBS             2602..2610
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in <i>E. coli</i>; complementary 
                     to a region in the 3' end of the 16S rRNA (<a 
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/7528374"" 
                     title=""https://www.ncbi.nlm.nih.gov/pubmed/7528374"">Chen 
                     et al., 1994</a>)"
     misc_feature    complement(2618..2646)
                     /label=pFAB weak constitutive
     misc_feature    2844..2922
                     /label=pJEX-D Promoter
     misc_feature    2899..2922
                     /label=EilO operator
     primer_bind     complement(2915..2957)
                     /label=oBA2269_pJEX-SC101Gib_R
     misc_feature    2945..2950
                     /label=RBS
     primer_bind     2955..3006
                     /label=oBA3004_pJEX_ECOR31_mCpol_STANDARD_F
     misc_feature    2956..2957
                     /label=2>3
     misc_feature    2960..4381
                     /label=ECOR31_mCpol_Native_STANDARD
     primer_bind     complement(3223..3263)
                     /label=oBA3067__ECOR31promoter_entry_GG_R
     CDS             3260..3619
                     /codon_start=1
                     /transl_table=0
                     /label=mCpol-like
                     /translation="MYIAIDGDDVGRKITSSYLSNSEERLTYISNKLNDTTKKISKMLL
                     SNGFEIIFQAADGVTAKTDNEVNLNFVFDKIKSYSFDEITFSAGVGANLREAYVALLNS
                     KSNGKNMISIYKDIL"
     primer_bind     complement(3348..3389)
                     /label=oBA2350_ECOR31_mCpol_UNK_Native_Gib_p1_R
     primer_bind     3348..3389
                     /label=oBA2351_ECOR31_mCpol_UNK_Native_Gib_p2_F
     misc_feature    3370..3377
                     /label=Problem_region_ECOR31
     CDS             3629..4231
                     /codon_start=1
                     /transl_table=0
                     
                     /translation="MFRIIRFDWLLKTVSLISLFVTILSFWIFKTVLSSDLSLLKLLSI
                     SSLASIIINFFLLSSKISRLIWGTIRFFKKDIYPDLNGIWVGHIITETNDEFEVRAKIK
                     QALLVTKIEMHGPTVKSVTLEATPTKELDNNKLYYVYKSTPKNPSWSEYIGSTIFDVIE
                     SNNALQLSGRYYTDRKSVGRISIKRISLNTDSDISFY"
     primer_bind     complement(4346..4385)
                     /label=oBA3005_pJEX_ECOR31_mCpol_STANDARD_R
     primer_bind     4381..4412
                     /label=oBA3068__pJEX_entry_GG_F
     primer_bind     4382..4412
                     /label=oBA2261_express_common_F
     misc_feature    4382..4385
                     /label=4bp BsaI
     terminator      4389..4517
                     /label=BBa_B0015 Terminator
     primer_bind     4389..4420
                     /label=oBFC0174
     primer_bind     4389..4412
                     /label=oBFC1711
     primer_bind     4397..4418
                     /label=oBFC0177
     primer_bind     complement(4425..4468)
                     /label=oBFC0177
     primer_bind     complement(4449..4475)
                     /label=oBFC1189
     primer_bind     complement(4491..4517)
                     /label=oBFC0696
     primer_bind     complement(4494..4518)
                     /label=oBFC0055
     primer_bind     complement(4494..4518)
                     /label=oBFC0806
     primer_bind     complement(4494..4517)
                     /label=oBFC0422
     primer_bind     complement(4494..4517)
                     /label=oBFC0426
     primer_bind     complement(4494..4517)
                     /label=oBFC0488
     primer_bind     complement(4494..4517)
                     /label=oBFC0490
     primer_bind     complement(4495..4538)
                     /label=oBA1653_RFP924_droupout_Gib_R
     primer_bind     complement(4496..4544)
                     /label=oBA1708_RFPdropout_part4_gib_R
     primer_bind     complement(4496..4532)
                     /label=oBFC1287
     primer_bind     complement(4496..4518)
                     /label=oBFC0309
     primer_bind     complement(4496..4518)
                     /label=oBFC1335
     primer_bind     complement(4496..4518)
                     /label=oBFC0785
     primer_bind     complement(4496..4518)
                     /label=oBFC0356
     primer_bind     complement(4496..4517)
                     /label=oBFC0692
     primer_bind     complement(4496..4511)
                     /label=oBFC1196
     primer_bind     4498..4541
                     /label=oBA1650_pBFC0924_Gib_F
     primer_bind     complement(4500..4518)
                     /label=oBFC1204
     primer_bind     complement(4500..4518)
                     /label=oBFC0738
     primer_bind     complement(4510..4538)
                     /label=oBFC1476
     primer_bind     4518..4567
                     /label=oBA1710_SC101_part4_gib_R
     misc_feature    4518..4544
                     /label=Pt4_common_region_6>4
     primer_bind     4518..4544
                     /label=oBA2199_pBA1169_BB_Gib_F
     primer_bind     complement(4523..4545)
                     /label=oBFC1462
     primer_bind     4524..4544
                     /label=oBA1739_pt4_commonGib_F
     misc_feature    4526..4529
                     /label=6>4 OH
     misc_feature    complement(4530..4533)
                     /label=SMRT Gate OH Right
     misc_feature    complement(4538..4544)
                     /label=AarI
     primer_bind     4543..4569
                     /label=oBFC0802
     misc_feature    4545..6807
                     /label=SC101_part
     primer_bind     4545..4569
                     /label=oBFC0693
     rep_origin      complement(4584..6052)
                     /direction=LEFT
                     /label=SC101
                     /note="/snapgene_direction=LEFT"
     primer_bind     complement(4745..4768)
                     /label=oBFC0727
     CDS             complement(5102..6052)
                     /codon_start=1
                     /transl_table=0
                     /gene="<i>rep101</i>"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg
       61 ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc
      121 ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg
      181 ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt taatcactat
      241 accaattgag atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta
      301 tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctata ccctctgtaa
      361 attccgctag acctttgtgt gttttttttg tttatattca agtggttata atttatagaa
      421 taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac
      481 tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa
      541 acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga
      601 atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag
      661 ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta
      721 tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg
      781 cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg
      841 ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc
      901 taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtccctgtg
      961 tcagatttct tactttattc aggcgattac gtgttacaac caattaacca attctgatta
     1021 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1081 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1141 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1201 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1261 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     1321 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1381 ctgagcgagg cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1441 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1501 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     1561 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     1621 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     1681 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     1741 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     1801 ggagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     1861 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     1921 attttgagac acaacgtggc tttgttgaat aaatcgaact tttgctgagt tgaaggatca
     1981 gcacctgccc gaaccagaga cttatctcag gacgtcttac gaaaataact caagctgaat
     2041 aacgtgctgc agatggcggg tgaatgcggc atcgtctacc tccggcattc ccagcacgta
     2101 aatcccgtcc agtccacaga ccagcgaaat cagccgccag gcgatatttt cggcgctatc
     2161 gcgcagggta aattcgccag cggcatggcc cgcgcgaatg atcctgaccg cttcgtcatg
     2221 ccacaggttc atggtcagca ggtacgcgct tttgatttcc ggatcgctgt cggccagcag
     2281 ctgcgcctgg cgccacaggc ggatataggg ctccagccgt ccgtcctcgc tgccgagggc
     2341 ggaaaacagc tgttcacgcc agccggcggt gcgcgaaagg cgctgcagat ccatcatctc
     2401 gcggatcacg cggataaacg cctgggattt tagctcgccg gaggaggtga aatggtggtg
     2461 gacctggcct gcggcgacgc ccgcggcggt ggcgatgttg cggaccgtca taccggtaaa
     2521 cccttgatcc agcgcgacgc gcatggcggc ctgcatgatg gtttcccgac gttcttcgcg
     2581 attcagatag cccatattgt cacctcctta tccacacatt atacgagccg gatgattaat
     2641 ggggaacctt accagagggc gccccagctg gcaattcctc ttaagtaagt aagagtatac
     2701 gtatatcggc taataacgta ttaaggcgct tcggcgcctt tttttatggg ggtattttca
     2761 tcccaatcca cacgtccaac gcacagcaaa caccacgtcg accctatcag ctgcgtgctt
     2821 tctatgagtc gttgctgcat aacaaaaaac tgcaaaaaat agtttgacag gacacgtgtc
     2881 caactttaag atgtacccaa agttggacac gtgtccaact ttgaattcaa aagatctttt
     2941 aagaaggaga tatactgatt gagaaatctt tagtgtaaag atttaataat tatcctttaa
     3001 gtgtagatta caagcagaac aattgaatta atcgccacat tgaaaatata aaccctcaac
     3061 acacttgatg tatttaacaa gacataggat taaacccatt aaaattaaaa aaatttaaaa
     3121 tacaaacctc gttttggccg ggtataaata caatttaaac cgcaacgttt ttaagtaaat
     3181 aaattcattt gcaactaagc attcagggtt gatataatat aaccacttag ataataatcc
     3241 atgttagttt ggagttaata tgtacatagc gatagatggc gatgatgtgg ggagaaaaat
     3301 aacctcatca tacttatcca actcagaaga gaggttgaca tacataagca acaaattaaa
     3361 tgataccaca aaaaaaatat ctaaaatgct tttatccaat ggatttgaga taattttcca
     3421 agccgcagat ggtgttacag cgaaaacaga caacgaggtc aatcttaact ttgtttttga
     3481 taagataaaa tcctattcct ttgacgaaat tacattttca gctggtgtgg gagcgaatct
     3541 aagagaagcc tatgttgcac tgttaaattc taaaagcaac ggtaagaata tgatttctat
     3601 ttacaaagac atattataat ggtgcgaaat gtttagaata ataagatttg actggttgtt
     3661 aaaaacggta tcactaatct ctttgtttgt cacaatactt tcattttgga tctttaagac
     3721 agttctgtct tctgacttaa gcctgctcaa attgttatca atatcatcat tagcatccat
     3781 tatcataaac ttctttctcc tttcatcaaa gatatcaaga ctaatatggg gaacaataag
     3841 gtttttcaag aaagacatat atcctgactt aaatggaatt tgggttggac atataatcac
     3901 agaaacaaat gatgaatttg aagtccgcgc aaaaataaag caagcactcc ttgtaacaaa
     3961 gatagaaatg cacggcccca cagttaaatc tgttacgcta gaagcaacgc caactaagga
     4021 actggataac aacaaattgt attatgtata taaatcaacc cctaaaaacc ctagttggag
     4081 cgagtatatt ggctcgacta tattcgacgt aattgaaagt aacaatgcgc tccaactttc
     4141 cggtcgatat tatacagaca gaaaatccgt ggggagaata tcaataaaaa gaatatcact
     4201 taacactgac tccgatatat ctttttatta actttcggaa tgtttaatta ccacaatcag
     4261 ccatgttcgt tcttttaaaa tagaaggata attcgcgcta ctgatatatt aatcatggaa
     4321 aaactccttt cagtagccat tttccctaga aaaatacccc tgcatacctc atatactcca
     4381 taggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     4441 tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg
     4501 gcctttctgc gtttatagaa gaccaaggag gactttggca ggtgctctcc tgagtaggac
     4561 aaatccgccg ccctagacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt
     4621 gttgctagtt tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagca
     4681 ctatttcttc cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt
     4741 tgtcggcagc tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg
     4801 ttgagctgta acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact
     4861 ggtttcacct gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc
     4921 atggtgaaca gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt
     4981 tacaccgttt tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt
     5041 tgttctactt ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac
     5101 ctcagatcct tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga
     5161 gccatgagaa cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct
     5221 caaaactggt gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt
     5281 tatgtaggta ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct
     5341 ggttgttctc aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca
     5401 acgtatcagt cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta
     5461 aatctttact tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat
     5521 tttcaagcat taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag
     5581 ttttcttttg tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt
     5641 caaaagactt aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag
     5701 gcaatatctc ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg
     5761 tccactggaa aatctcaaag cctttaacca aaggattcct gatttccaca gttctcgtca
     5821 tcagctctct ggttgcttta gctaata
//
