LOCUS       Exported               10703 bp ds-DNA     circular SYN 22-JUL-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10703)
  AUTHORS   The Weissman lab
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 6, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    874..936
                     /label=5'ITR
     misc_feature    complement(1400..1993)
                     /label=WPRE
                     /label=WPRE; Woodchuck post-transcriptional
     primer_bind     complement(1893..1912)
                     /label=HKO083
     CDS             complement(2016..2615)
                     /label=Puro (-)
     promoter        complement(2628..3130)
                     /label=PGK (-)
     misc_feature    3137..8069
                     /label=UQCRQ
     misc_feature    3961
                     /label=G>A in clone
     misc_feature    4336
                     /label=G>A in clone
     misc_feature    4418
                     /label=G>A in clone
     misc_feature    4877..4922
                     /label=5'UTR
     misc_feature    4923..5103
                     /label=5UTRintron
     misc_feature    4942
                     /label=T>C in clone
     misc_feature    4954
                     /label=C>G in clone
     misc_feature    5104..5116
                     /label=5'UTR
     misc_feature    5117..5270
                     /label=exon1
     misc_feature    5158
                     /label=syn G>C in clone
     misc_feature    5162..5176
                     /label=recoding region
     misc_feature    5179
                     /label=syn G>A in clone
     misc_feature    5271..5722
                     /label=intron
     misc_feature    5723..5817
                     /label=exon2
     misc_feature    5818..7081
                     /label=3'UTR
     misc_feature    6991
                     /label=del G right after in clone
     misc_feature    7009
                     /label=A>G in clone
     misc_feature    7754
                     /label=G>A in clone
     misc_feature    8157..8191
                     /label=3'ITR
     primer_bind     complement(8682..8698)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8706..8722
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(8730..8760)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8775..8796
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(9084..9672)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9843..10703)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc agggtttatt acagggacag cagagatcca
     1381 gtttggttaa ttagctagcg atgcggggag gcggcccaaa gggagatccg actcgtctga
     1441 gggcgaaggc gaagacgcgg aagaggccgc agagccggca gcaggccgcg ggaaggaagg
     1501 tccgctggat tgagggccga agggacgtag cagaaggacg tcccgcgcag aatccaggtg
     1561 gcaacacagg cgagcagcca aggaaaggac gatgatttcc ccgacaacac cacggaattg
     1621 tcagtgccca acagccgagc ccctgtccag cagcgggcaa ggcaggcggc gatgagttcc
     1681 gccgtggcaa tagggagggg gaaagcgaaa gtcccggaaa ggagctgaca ggtggtggca
     1741 atgccccaac cagtgggggt tgcgtcagca aacacagtgc acaccacgcc acgttgcctg
     1801 acaacgggcc acaactcctc ataaagagac agcaaccagg atttatacaa ggaggagaaa
     1861 atgaaagcca tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat
     1921 agcgtaaaag gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc
     1981 agaggttgat taccgataag cttgatatcg aattcttaac tggcaccggg cttgcgggtc
     2041 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     2101 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     2161 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     2221 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     2281 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     2341 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     2401 acggcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     2461 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccggatcgac ggtgtggcgc
     2521 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggccctggg gacgtcgtcg
     2581 cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg tggtaccccc tggggagaga
     2641 ggtcggtgat tcggtcaacg agggagccga ctgccgacgt gcgctccgga ggcttgcaga
     2701 atgcggaaca ccgcgcgggc aggaacaggg cccacactac cgccccacac cccgcctccc
     2761 gcaccgcccc ttcccggccg ctgctctcgg cgcgccctgc tgagcagccg ctattggcca
     2821 cagcccatcg cggtcggcgc gctgccattg ctccctggcg ctgtccgtct gcgagggtac
     2881 tagtgagacg tgcggcttcc gtttgtcacg tccggcacgc cgcgaaccgc aaggaacctt
     2941 cccgacttag gggcggagca ggaagcgtcg ccggggggcc cacaagggta gcggcgaaga
     3001 tccgggtgac gctgcgaacg gacgtgaaga atgtgcgaga cccagggtcg gcgccgctgc
     3061 gtttcccgga accacgccca gagcagccgc gtccctgcgc aaacccaggg ctgccttgga
     3121 aaaggcgcaa acgcgttgga ttaagatgtt cgctaggtgc ttgctgaaac ttctaaggca
     3181 tgtggaaaag ctaaccaatt attcatgacg atactcagat ctgctggtat gcaacttatg
     3241 gtattaggtt tactcttcct ctcaatttat cctcacggcc ttttccactt ccaggtgttt
     3301 acaattgtaa ttattcgctg catgttaatt tgcgaattta caaatttggg tgagaaagtc
     3361 acaattggca tagaagttcc agaaagcaga aaaccattag gtctctcagt cctcccaggt
     3421 ggctgggtac agaatacgta cttggagtac tggaattgta tttgcgatgt tcttaaaaga
     3481 ggacaccgcc tttaaccttg acttattttt aaatgtaaca tactgatgtc cttactttgg
     3541 tctgcacact catctataca tattttggtt gaaaggtttt gggatgtttg cagatgctat
     3601 aaaataaatg aacgttttat tattattcat ccaaatggtg gtagaatttg ccacaatatt
     3661 tctgtcctta cctgtatttc tgttcttgcc ttggagctga gaccaggcta gacattatgg
     3721 tttaaaaaaa tgaggcaaag gagcaccatg aaaagctgaa atgaggggat attcagcgcc
     3781 accccgaagg agactatgct ggaagagtca gcctacactc atagtgcaag atacgaagcc
     3841 agtttgtgca gtacctacat gagaaaacta aggtatgtct gcaaaataca agttgactgc
     3901 ctgccgagcc tccgtggtcc gtaagtagtg ttataaaaca cagccaactg ttgaatgaga
     3961 acttagtcct agataaaata tggccagcaa ttccattaac aaaacactga caagacacct
     4021 cagggcgagc tgtacattga caacagcatc gtcaagcaga atgcacttca gggaaaacat
     4081 aataaggcct ctcacccgac tttccttgct tctcacgtgg ggagaagaaa gaactggggc
     4141 atcgtgattc cccccacccc acgcaaacca tctggggagg acgctggctg ctgtcgaggc
     4201 ggcctcagca ttcatcaatt gacgattatt tgattatttt tgtggctgaa actgacgaaa
     4261 cgtttgtatt atctcttggg aacgttaaca gcctgccagg ggactctaaa acagagtcca
     4321 agagaagcct tcgatatgtg tccctgattt tggtgctggg gtcgctcatg gcctgaaaat
     4381 aacgaagccc cctttgagct actggccaag gccaaataaa acctcgtgct cctctgctgt
     4441 ggggcggccc ggcctcgggg ctcagaggcg ctctgtctac agctgggaag actcgcaggc
     4501 cccaagctcc tggccagcgc tgccccggag gtcggcaggc cccttcctcg tcaccttttt
     4561 gttccctccc ccgcctcccg cattcggccg cttcctgact gggattccac agaaaagccg
     4621 agggctgagg agaagtgtga gcgcctccgc ctgtccactg tcccccaaag tcagttcaat
     4681 ccccgacgtc ctccgctagg ctccacccca ccggcccggg cagggcctcc aaggcacctc
     4741 ccacctacgg gtcacccagt cagcccactt ctttctggga caaaggcgtc atcccttaga
     4801 gacagtagga aaatggtatc tcccggaagt tacctcacga cctccaagag cggcttccaa
     4861 ccttgccgga aatgacgaac gagtcaaccg gatcggtgac tgtggagggc gagctgagcc
     4921 ctgtgcgtga gtggggtctg gctgtgcagt gttggtggac cctgggaggc taggggcgcc
     4981 ccgctgggct gggaaaggat aaggagtgca ggggcaggag tctggggttg gggatggacc
     5041 cccgcgggga ctgcggcgct tcgcgaaagc gagccaagcg cctgtccacc ctcggtcctg
     5101 cagggccgcc gccacaatgg gccgcgagtt tgggaatctg acgcggatgc ggcatgtcat
     5161 ctcctactcc ttgagtccat tcgagcagcg cgcctatccg cacgtcttca ctaaaggaat
     5221 ccccaatgtt ctgcgccgca ttcgggagtc tttctttcgc gtggtgccgc gtgagtgcct
     5281 tgggcccgcg ggagcgggag gctggacccc agcagcagca gcagtcactg cgcctcccct
     5341 ctgagcgctg gcggccgggc aggcgctctg taacttgccg tacacacctt tctcattgaa
     5401 tattcctgac cgcccttaag ctggtcatat tatcctcccc atcttacacg gggaaaactg
     5461 agctacagaa ctgaaatgac tggcccaagg tcactcgggt gattccagca tgttcctctg
     5521 aggtcacctt tatttagttc ttaccagtgg tgaaatgctt tgtgttaaac gtgcagcaga
     5581 tagtgatgat agttgcatgg ccttggatat gctaagaact actgaagtgt atgctttaaa
     5641 gaagacaaga ggactgagac cttttttttt cttctatatg tcttatatgt gtgctctctg
     5701 tttactctct taatttttaa agagtttgta gtgttttatc ttatctacac atgggggact
     5761 gaagagttcg agagatccaa gaggaagaat ccagctgcct atgaaaatga caaatgagca
     5821 acgcatccgg atgacggttc cctgtctctg aaagaccttt ctctggaaga ggagtctgca
     5881 ttgtagtgtc tcaaagacac aataaacttc ctatggtctg cactgttgtg atattacatt
     5941 tttgtgagta gaatcctgtg tgaccactaa tattcaagtt catgaagcgc ccctcctcag
     6001 catgagtctt gaagttttgc ctggtctcgg gttttggtga aagagcaggg ctcccccttt
     6061 gttccattat ctacaagaaa gtagctgcag gtactcctgc atctaaccat ttgtgggtga
     6121 catgggctat ggcgagtgct ggttcttggc cctctgctcc catagcaaca ataaagtcta
     6181 ctaactatat tgtaattagt atgtgtcagg gacgctttta ttttttattt tttgagatgg
     6241 agtcttgctc tctcacccag gctggagtgc agtggtgtga tctcggctca ctgccatctg
     6301 tgcctcctgg gttcaaacga ttctcctgtc tcagcctccg agtagctggg attacaggtg
     6361 caggacacca cacctggcta attttttgta tttttagtag agacaggttt caccatgttg
     6421 gccaggctgg tctggaactc ctgacctcag atgatctgtg cgcctcagcc tcccaaagtg
     6481 ctgggattac aggcgtaagc catggtgctc tgggcctcaa atactcttct tagctacaca
     6541 aatgaactgt taaacctcac aagcaccttt aacgtatcct catttggcag atgaggaaat
     6601 gggcacagag agattaagta atttgccaga aattatacag cagtgaatgg tggagtcttg
     6661 cactaaagtg atgctcttaa aactatgctc tattgcaggg gtgtccaatc ttttagcttc
     6721 cctgggccac acataaaata cgctaacatt acaatagctg ataagcttta aaaaatggca
     6781 aaaaaaaccc tcagcgtttt agaaagttta caaatttttg ttgggtcgga ttcaaagcca
     6841 tcctgggctg cttgctgccc aggactgcgg ttggacaagc ttgctctact gcctttcctc
     6901 ctaaggcagg ccttagcctg aggggatatc agctataaaa cggatgaggt gatctcttag
     6961 cctccacagc agttttctgt ttctgtgcca gaggccaaca atgttttcga ccatatgtgg
     7021 ttcataaagc ctcaaagatt tactctggcc ctttatgaat aaagtttacc cctgatctaa
     7081 agcctttaaa ttatcttttt ggtattaaag ataccatttt cccttcgccc accgagagat
     7141 ggtgggggta ggtgtggttg gcagttttat gcctctagtt ccttagagag gcttctctct
     7201 aagcttcggg acatggatct cacctgtggt agcaggtatg tggaatatga tgagttgtca
     7261 tagctcagtc agtacagtat atgctatggg atctgagatc cgtttttaga aactcttcta
     7321 aggactcttc ctgctagcag caactttgat tctgggtcag actgtgtttt tttttttttt
     7381 tttttgagat gggagtcttg ctctgtcacc aggctggagt ttagtggcgc aatctcagct
     7441 tactgcaacc tccgcttccc gggctcaagt gattctcctg cctcagcctc ccaaatagct
     7501 gggactacag gcgcatacca acacacccag ctaatttttg tatttttatt ttttttttta
     7561 gtagagacgg agtttcacca tgttggccag gatggtcttg atctcttgac ctcgtgatcc
     7621 acctgcctcg acctcccaaa gtgctgggat tacaggtgtg agccaccgtg cctggccaga
     7681 ctgtatcttt attatcctaa ttactgctat tgccttttaa gaatcaaaag ttaagatact
     7741 tcaaaacatg gccagataca gtggctcaca cctgtaatcc cagcactttg ggaggctgaa
     7801 gtggatggat cacttgaggt caggagtttg agaccagcct ggccaacatg gtgaaaccct
     7861 gtctctacta aaaatataaa aattagctga gtgtggtggc acacgcctat aatcccagct
     7921 actcaggagg ctgaggcaga aacgcttgaa cctgggaggt ggaggttgca gtgagccaag
     7981 attgtgccac tgcactccag cctgggtgac agagcgagac ttcaaaacaa aacctaaaag
     8041 ataaactaca ttgtgaggca ggatacgggg tttaaaccaa gcggcgactg agatgtccta
     8101 aatgcacagc gacggattcg cgctatttag aaagagagag caatatttca agaatgcatg
     8161 cgtcaatttt acgcagacta tctttctagg gttaatctag ctgcatcagg atcatatcgt
     8221 cgggtctttt ttccggctca gtcatcgccc aagctggcgc tatctgggca tcggggagga
     8281 agaagcccgt gccttttccc gcgaggttga agcggcatgg aaagagtttg ccgaggatga
     8341 ctgctgctgc attgacgttg agcgaaaacg cacgtttacc atgatgattc gggaaggtgt
     8401 ggccatgcac gcctttaacg gtgaactgtt cgttcaggcc acctgggata ccagttcgtc
     8461 gcggcttttc cggacacagt tccggatggt cagcccgaag cgcatcagca acccgaacaa
     8521 taccggcgac agccggaact gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct
     8581 gggatattac gtcagcgagg acgggtatcc tggctggatg ccgcagaaat ggacatggat
     8641 accccgtgag ttacccggcg ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     8701 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     8761 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     8821 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     8881 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     8941 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     9001 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     9061 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     9121 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     9181 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     9241 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     9301 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     9361 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     9421 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     9481 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     9541 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     9601 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     9661 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     9721 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     9781 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     9841 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     9901 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     9961 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
    10021 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
    10081 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
    10141 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
    10201 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
    10261 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
    10321 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
    10381 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
    10441 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
    10501 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
    10561 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
    10621 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
    10681 acacggaaat gttgaatact cat
//
