LOCUS       Exported                5101 bp ds-DNA     circular SYN 19-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5101)
  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: EC100D(TM) pir+SnapGene UUID: 
            40a8c859-f016-4ee7-b1f4-5772e5ceb948Sequence Document Type: UNAOther
            Notes: CreatedBy: Brady CressImported FromPath: 
            /Users/benjaminadler/Library/CloudStorage/GoogleDrive-ba14@berkeley.
            edu/My Drive/BA_Doudna_Lab/BA_DNA_DB/BA_SnapGene/DNA 
            Files/CoexpressionFilename: pBA1326_pJEX-mRFP_SC101Kan.dna
FEATURES             Location/Qualifiers
     source          1..5101
                     /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
     primer_bind     2928..2981
                     /label=oBA2262_RFP_CDS_pJEXGib_F
     misc_feature    2945..2950
                     /label=RBS
     misc_feature    2956..2959
                     /label=2>3
     misc_feature    2958..3635
                     /label=mRFP
                     /label=RFP
     misc_feature    complement(2959..2989)
                     /label=RFP_CDS_13a_1
     misc_feature    3000..3027
                     /label=Cpf1 target 2
     misc_feature    3040..3070
                     /label=C PAM 2
     misc_feature    3043..3073
                     /label=A PAM 2
     misc_feature    3044..3074
                     /label=G PAM 2
     misc_feature    3046..3076
                     /label=T PAM 2
     misc_feature    complement(3057..3087)
                     /label=Target 2
     misc_feature    complement(3058..3088)
                     /label=target 3
     misc_feature    complement(3059..3089)
                     /label=Target  1
     misc_feature    3061..3088
                     /label=Cpf1 target 1
     misc_feature    complement(3111..3141)
                     /label=target 4 (G PAM)
     misc_feature    3145..3175
                     /label=T PAM 0
     misc_feature    3148..3178
                     /label=A PAM 0
     misc_feature    3153..3183
                     /label=C PAM 0
     misc_feature    complement(3154..3184)
                     /label=target 6 (C PAM)
     misc_feature    3154..3184
                     /label=G PAM 0
     misc_feature    complement(3155..3173)
                     /label=RFP RT primer 1
     misc_feature    complement(3187..3217)
                     /label=target 5 (G PAM)
     misc_feature    3222..3252
                     /label=T PAM 1
     misc_feature    3223..3253
                     /label=C PAM 1
     misc_feature    3226..3256
                     /label=A PAM 1
     misc_feature    3227..3257
                     /label=G PAM 1
     misc_feature    complement(3255..3270)
                     /label=RFP RT Primer 2
     misc_feature    complement(3257..3270)
                     /label=RT pr 1
     misc_feature    complement(3257..3270)
                     /label=RT p1
     misc_feature    3295..3325
                     /label=A PAM 3
     misc_feature    3296..3326
                     /label=C PAM 3
     misc_feature    3297..3327
                     /label=T PAM 3
     misc_feature    3298..3328
                     /label=G PAM 3
     misc_feature    complement(3355..3370)
                     /label=RT p2
     misc_feature    complement(3455..3470)
                     /label=RT p3
     primer_bind     complement(3615..3665)
                     /label=oBA2263_RFP_CDS_Gib_R
     primer_bind     3636..3666
                     /label=oBA2261_express_common_F
     terminator      3643..3771
                     /label=BBa_B0015 Terminator
     primer_bind     3643..3674
                     /label=oBFC0174
     primer_bind     3643..3666
                     /label=oBFC1711
     primer_bind     3651..3672
                     /label=oBFC0177
     primer_bind     complement(3679..3722)
                     /label=oBFC0177
     primer_bind     complement(3703..3729)
                     /label=oBFC1189
     primer_bind     complement(3745..3771)
                     /label=oBFC0696
     primer_bind     complement(3748..3772)
                     /label=oBFC0055
     primer_bind     complement(3748..3772)
                     /label=oBFC0806
     primer_bind     complement(3748..3771)
                     /label=oBFC0422
     primer_bind     complement(3748..3771)
                     /label=oBFC0426
     primer_bind     complement(3748..3771)
                     /label=oBFC0488
     primer_bind     complement(3748..3771)
                     /label=oBFC0490
     primer_bind     complement(3749..3792)
                     /label=oBA1653_RFP924_droupout_Gib_R
     primer_bind     complement(3750..3798)
                     /label=oBA1708_RFPdropout_part4_gib_R
     primer_bind     complement(3750..3786)
                     /label=oBFC1287
     primer_bind     complement(3750..3772)
                     /label=oBFC0309
     primer_bind     complement(3750..3772)
                     /label=oBFC1335
     primer_bind     complement(3750..3772)
                     /label=oBFC0785
     primer_bind     complement(3750..3772)
                     /label=oBFC0356
     primer_bind     complement(3750..3771)
                     /label=oBFC0692
     primer_bind     complement(3750..3765)
                     /label=oBFC1196
     primer_bind     3752..3795
                     /label=oBA1650_pBFC0924_Gib_F
     primer_bind     complement(3754..3772)
                     /label=oBFC1204
     primer_bind     complement(3754..3772)
                     /label=oBFC0738
     primer_bind     complement(3764..3792)
                     /label=oBFC1476
     primer_bind     3772..3821
                     /label=oBA1710_SC101_part4_gib_R
     misc_feature    3772..3798
                     /label=Pt4_common_region_6>4
     primer_bind     complement(3777..3799)
                     /label=oBFC1462
     misc_feature    3780..3783
                     /label=6>4 OH
     misc_feature    complement(3784..3787)
                     /label=SMRT Gate OH Right
     misc_feature    complement(3792..3798)
                     /label=AarI
     primer_bind     3797..3823
                     /label=oBFC0802
     misc_feature    3799..6061
                     /label=SC101_part
     primer_bind     3799..3823
                     /label=oBFC0693
     rep_origin      complement(3838..5306)
                     /direction=LEFT
                     /label=SC101
                     /note="/snapgene_direction=LEFT"
     primer_bind     complement(3999..4022)
                     /label=oBFC0727
     CDS             complement(4356..5306)
                     /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 tattctgatg gcgagtagcg aagacgttat caaagagttc atgcgtttca
     3001 aagttcgtat ggaaggttcc gttaacggtc acgagttcga aatcgaaggt gaaggtgaag
     3061 gtcgtccgta cgaaggtacc cagaccgcta aactgaaagt taccaaaggt ggtccgctgc
     3121 cgttcgcttg ggacatcctg tccccgcagt tccagtacgg ttccaaagct tacgttaaac
     3181 acccggctga catcccggac tacctgaaac tgtccttccc ggaaggtttc aaatgggaac
     3241 gtgttatgaa cttcgaagac ggtggtgttg ttaccgttac ccaggactcc tccctgcaag
     3301 acggtgagtt catctacaaa gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg
     3361 ttatgcagaa aaaaaccatg ggttgggaag cttccaccga acgtatgtac ccggaagacg
     3421 gtgctctgaa aggtgaaatc aaaatgcgtc tgaaactgaa agacggtggt cactacgacg
     3481 ctgaagttaa aaccacctac atggctaaaa aaccggttca gctgccgggt gcttacaaaa
     3541 ccgacatcaa actggacatc acctcccaca acgaagacta caccatcgtt gaacagtacg
     3601 aacgtgctga aggtcgtcac tccaccggtg cttaaaggat ctccaggcat caaataaaac
     3661 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     3721 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat agaagaccaa
     3781 ggaggacttt ggcaggtgct ctcctgagta ggacaaatcc gccgccctag acctagggta
     3841 cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct agtttgttat cagaatcgca
     3901 gatccggctt cagccggttt gccggctgaa agcactattt cttccagaat tgccatgatt
     3961 ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca
     4021 gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
     4081 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
     4141 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa
     4201 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
     4261 acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg
     4321 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
     4381 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
     4441 tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
     4501 gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg
     4561 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
     4621 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
     4681 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
     4741 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
     4801 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
     4861 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
     4921 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
     4981 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
     5041 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
     5101 a
//
