LOCUS       Exported               10070 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 10070)
  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..10070
                     /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..7436
                     /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..4935
                     /label=5'UTR
     misc_feature    4936..5184
                     /label=UQCRQ CDS
     misc_feature    4936..5089
                     /label=exon1
     misc_feature    4981..4995
                     /label=recoding region
     misc_feature    5090..5184
                     /label=exon2
     misc_feature    5185..6448
                     /label=3'UTR
     misc_feature    6358
                     /label=del G right after in clone
     misc_feature    6376
                     /label=A>G in clone
     misc_feature    7121
                     /label=G>A in clone
     misc_feature    7524..7558
                     /label=3'ITR
     primer_bind     complement(8049..8065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8073..8089
                     /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(8097..8127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8142..8163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8451..9039)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9210..10070)
                     /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 ctggccgccg ccacaatggg ccgcgagttt gggaatctga cgcggatgcg gcatgtgatc
     4981 tcctactcct tgagtccgtt cgagcagcgc gcctatccgc acgtcttcac taaaggaatc
     5041 cccaatgttc tgcgccgcat tcgggagtct ttctttcgcg tggtgccgca gtttgtagtg
     5101 ttttatctta tctacacatg ggggactgaa gagttcgaga gatccaagag gaagaatcca
     5161 gctgcctatg aaaatgacaa atgagcaacg catccggatg acggttccct gtctctgaaa
     5221 gacctttctc tggaagagga gtctgcattg tagtgtctca aagacacaat aaacttccta
     5281 tggtctgcac tgttgtgata ttacattttt gtgagtagaa tcctgtgtga ccactaatat
     5341 tcaagttcat gaagcgcccc tcctcagcat gagtcttgaa gttttgcctg gtctcgggtt
     5401 ttggtgaaag agcagggctc cccctttgtt ccattatcta caagaaagta gctgcaggta
     5461 ctcctgcatc taaccatttg tgggtgacat gggctatggc gagtgctggt tcttggccct
     5521 ctgctcccat agcaacaata aagtctacta actatattgt aattagtatg tgtcagggac
     5581 gcttttattt tttatttttt gagatggagt cttgctctct cacccaggct ggagtgcagt
     5641 ggtgtgatct cggctcactg ccatctgtgc ctcctgggtt caaacgattc tcctgtctca
     5701 gcctccgagt agctgggatt acaggtgcag gacaccacac ctggctaatt ttttgtattt
     5761 ttagtagaga caggtttcac catgttggcc aggctggtct ggaactcctg acctcagatg
     5821 atctgtgcgc ctcagcctcc caaagtgctg ggattacagg cgtaagccat ggtgctctgg
     5881 gcctcaaata ctcttcttag ctacacaaat gaactgttaa acctcacaag cacctttaac
     5941 gtatcctcat ttggcagatg aggaaatggg cacagagaga ttaagtaatt tgccagaaat
     6001 tatacagcag tgaatggtgg agtcttgcac taaagtgatg ctcttaaaac tatgctctat
     6061 tgcaggggtg tccaatcttt tagcttccct gggccacaca taaaatacgc taacattaca
     6121 atagctgata agctttaaaa aatggcaaaa aaaaccctca gcgttttaga aagtttacaa
     6181 atttttgttg ggtcggattc aaagccatcc tgggctgctt gctgcccagg actgcggttg
     6241 gacaagcttg ctctactgcc tttcctccta aggcaggcct tagcctgagg ggatatcagc
     6301 tataaaacgg atgaggtgat ctcttagcct ccacagcagt tttctgtttc tgtgccagag
     6361 gccaacaatg ttttcgacca tatgtggttc ataaagcctc aaagatttac tctggccctt
     6421 tatgaataaa gtttacccct gatctaaagc ctttaaatta tctttttggt attaaagata
     6481 ccattttccc ttcgcccacc gagagatggt gggggtaggt gtggttggca gttttatgcc
     6541 tctagttcct tagagaggct tctctctaag cttcgggaca tggatctcac ctgtggtagc
     6601 aggtatgtgg aatatgatga gttgtcatag ctcagtcagt acagtatatg ctatgggatc
     6661 tgagatccgt ttttagaaac tcttctaagg actcttcctg ctagcagcaa ctttgattct
     6721 gggtcagact gtgttttttt tttttttttt ttgagatggg agtcttgctc tgtcaccagg
     6781 ctggagttta gtggcgcaat ctcagcttac tgcaacctcc gcttcccggg ctcaagtgat
     6841 tctcctgcct cagcctccca aatagctggg actacaggcg cataccaaca cacccagcta
     6901 atttttgtat ttttattttt ttttttagta gagacggagt ttcaccatgt tggccaggat
     6961 ggtcttgatc tcttgacctc gtgatccacc tgcctcgacc tcccaaagtg ctgggattac
     7021 aggtgtgagc caccgtgcct ggccagactg tatctttatt atcctaatta ctgctattgc
     7081 cttttaagaa tcaaaagtta agatacttca aaacatggcc agatacagtg gctcacacct
     7141 gtaatcccag cactttggga ggctgaagtg gatggatcac ttgaggtcag gagtttgaga
     7201 ccagcctggc caacatggtg aaaccctgtc tctactaaaa atataaaaat tagctgagtg
     7261 tggtggcaca cgcctataat cccagctact caggaggctg aggcagaaac gcttgaacct
     7321 gggaggtgga ggttgcagtg agccaagatt gtgccactgc actccagcct gggtgacaga
     7381 gcgagacttc aaaacaaaac ctaaaagata aactacattg tgaggcagga tacggggttt
     7441 aaaccaagcg gcgactgaga tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa
     7501 gagagagcaa tatttcaaga atgcatgcgt caattttacg cagactatct ttctagggtt
     7561 aatctagctg catcaggatc atatcgtcgg gtcttttttc cggctcagtc atcgcccaag
     7621 ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc ttttcccgcg aggttgaagc
     7681 ggcatggaaa gagtttgccg aggatgactg ctgctgcatt gacgttgagc gaaaacgcac
     7741 gtttaccatg atgattcggg aaggtgtggc catgcacgcc tttaacggtg aactgttcgt
     7801 tcaggccacc tgggatacca gttcgtcgcg gcttttccgg acacagttcc ggatggtcag
     7861 cccgaagcgc atcagcaacc cgaacaatac cggcgacagc cggaactgcc gtgccggtgt
     7921 gcagattaat gacagcggtg cggcgctggg atattacgtc agcgaggacg ggtatcctgg
     7981 ctggatgccg cagaaatgga catggatacc ccgtgagtta cccggcgggc gcgcttggcg
     8041 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     8101 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     8161 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     8221 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     8281 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     8341 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     8401 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8461 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8521 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8581 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8641 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8701 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8761 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8821 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8881 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8941 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     9001 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     9061 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     9121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     9181 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     9241 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     9301 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     9361 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9421 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9481 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9541 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9601 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9661 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9721 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9781 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9841 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9901 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9961 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    10021 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
//
