LOCUS       Exported                9388 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 9388)
  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..9388
                     /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..6754
                     /label=COX7A2
     misc_feature    4187..5186
                     /label=Promoter 1000bp upstream
     misc_feature    5187..5253
                     /label=5' UTR
     misc_feature    5254..5505
                     /label=CDS
     misc_feature    5304..5312
                     /label=mutations
     misc_feature    5506..5648
                     /label=3' UTR
     misc_feature    5649..6648
                     /label=1000 bp downstream
     misc_feature    6842..6876
                     /label=3'ITR
     primer_bind     complement(7367..7383)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7391..7407
                     /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(7415..7445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7460..7481
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7769..8357)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8528..9388)
                     /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 acgcgttggt gagggacgga ttaactatgt taaatgctgc taatagatgg
     3181 agaaggtgaa aattgagatt gacagtttat ttagcaacct ggaagctatt ggtgattcta
     3241 aacaagagca gtaaaagcct gcccaaaatg ggatcaaaaa aagattagga ggaggagaag
     3301 gagagacaag tataaagaaa tctataggat ggtaacggta atcaaaggag tttgttcgtt
     3361 tgtttttaat ttgggaatgt gttaatagat ggaatgatcc agtaaagggg gaaattgtac
     3421 tgaataacat gcatacaaga atctatgtct actttaggat ctatttaatg tgaattcatg
     3481 gctttttttt tttttttgat atgaagtctc actctgtcgc ccaggctgga gtgcagtgga
     3541 gcgatcttgg ctcactgcaa gctccacctt ctgggttcac gccattctcc tgcctcaacc
     3601 tcctgagtag ctgggactac aggcgcccgc caccacgccc gccttatttt ttgtattttt
     3661 agtagagaca gggtttcacg gctttagcca agatggtctg gatctcctga cctcatgatc
     3721 cgcccgcctc ggcctcccaa agtactggga ttacaggcgt gagccactgc gcctggcctc
     3781 atggctcttt ttggacaggg tcataagctt cttagatctg tcacccaatg taattaattg
     3841 atgtattggt taatactgtc tttagcagtg ctaggggatt ggtaagaaat ccacatcatc
     3901 tctgcgcttg ggaaatttac aatctagagt tggctaagtg gagaaaacat aagagagcac
     3961 atacaactaa caaattcaga ttatggacaa tgtacttaaa aaacccccac gccctagcta
     4021 ttggcacgtg gtaattttgc ccagagtttt ggttatatgc ataatccaga ggggtgtaga
     4081 aaaccaggtc ctttcagtac caatctaagg aagacatgaa ttggggtgtt tataaacctt
     4141 ggtgaggaag agaattttaa ctgggactgg taacttggtg ggcccattga catacagtag
     4201 attgtgaaga agaaaacccc aaggattata aggaatagtt ttaatattac taatttttat
     4261 cctttgctga aactatgggt ttcttatttg aaatggcaat caaataattt gtaagagttt
     4321 gtagaaagtg gaaagattat tgtaatacag ggtagatatt tgcatttgta catacttgag
     4381 ggtaagtaca agaaataaac acttgtaatg gggattactt aagcaaaatg gatgacaccg
     4441 acagcctttc tgtacaacaa ataattgctg tgcttctcta ctccagtaat gaatgtagtg
     4501 ccctgctgga tctgggacag caagactttt gagtccagat tcgccatccc atttcttgtt
     4561 ccctaacgtt tcctcagaag cgagcgagcc tcttccacaa tttggggaca ctgtggaaga
     4621 acttcttgct tggggagttc tgcagcagtc gaaggtacaa gaatataaaa actggtatct
     4681 cgacacagct cttacagcat attctcaaca caaggaacag taatgtgccg cagcaattcc
     4741 ttcggtggaa aaaactgact gtatccgcgt aagaggagta attctttttt gaattttatg
     4801 aggcttcaga gctgtatttc ggccaaccca gtctttgttc agtttctagc ggcttcaagg
     4861 ttgcccagtt ttacgccttc tcgctcaaaa cagacactag ttttctttga tagcctggtc
     4921 tcacattgac ttcatctaag cttattacat tcgtctcata aatcgtttgc tttttcccca
     4981 gccttagctc cagggaccag cctactgaac atcgaaaagg tcaggaacga gatcacggga
     5041 ctgagacggc gagccggcta gagcggatag agccgtactg ccgctctagt ttttcttttg
     5101 ctctctatgg ttaggagcgc aagcctctct tcggcccgga aagatttaag ttcgtgaatg
     5161 catacgcaag actcggaggt agttccggtt ccggcgtggc cattttcgtt ggtggtgttc
     5221 agttgtggcg gttgctggtc agtaacagcc aagatgctgc ggaatctgct ggctcttcgt
     5281 cagattgggc agaggacgat aagaacagca agccgcaggc attttaaaaa taaagttccg
     5341 gagaagcaaa aactgttcca ggaggatgat gaaattccac tgtatctaaa gggtggggta
     5401 gctgatgccc tcctgtatag agccaccatg attcttacag ttggtggaac agcatatgcc
     5461 atatatgagc tggctgtggc ttcatttccc aagaagcagg agtgacttca gtcatcccag
     5521 caatcgcttg gttcagtttc attcagctct ctatggacca gtaatctgat aaataaccga
     5581 gctcttcttt ggggatcaat atttattgac ttgtagtaac tgccaccaat aaagcagtct
     5641 ttaccatgct ttgtctgatt ttatcacttt atgccaataa tttcaacttt tggtcagcct
     5701 tgtaaatctt aggtaaggat caaagaaatg attcttttgc cactaaactt gtgaaagcaa
     5761 cattattgtg atgttatttt agtgaaattt tacttctaac agctcaaaat tagtctcaac
     5821 ttctgtctca acttgccgtt tgaatgcata ttgctttttc cccctaccca ctccagtata
     5881 tcttggaata tgctagaacc cgggggtagt ggaagcagat tatctttggc tcctggggtc
     5941 tatatttctt tttttttttt tttttctcag ctatgtgaga gtatgcttaa atattggata
     6001 tatcccacag cttagaatta ggctttagta gaatggcttt gtaagatatt tcagtatttg
     6061 tttttttcta cactccgagt gggttcaggt aggtttctct catcaggaac caaccaaagc
     6121 aggacagttc ctgggctccc cagctatcat ctaatggacc cacaactgtt taacctaaac
     6181 agtagttagg tgcttaataa atggtagcca taataggctg ctcataatag gaatgtttca
     6241 aaaaggtcta tttagggaac acaaaatcca aatagataca gagagtaaga atcaaaaaga
     6301 attacgatgc tgctctaggt agtttgcttc agggtttgca gaacccttcc tgctttgaga
     6361 gaactatgca tgaagctgga aatttcatag acttcaagtt tgggactccc ttttcctggg
     6421 tcctgttgtg catccatgtg gagagtagca tcttgtgaac cacttcagga ctttccttct
     6481 atatatatcg agattggtaa ttcgtcaaga gatactgagt gcctactgtg tcctaactgg
     6541 ttttaggtac ctaggaatca ctggtgaaca aaagcagatt cttcccctca tatgttatag
     6601 aaaggggtgg taggaaagag agtttaaaaa taaattgcac agtaggttaa aagataagtt
     6661 ctatggaaaa agaccaaagg ggagaactca catgggtgtg gttggttggt tgcagtatta
     6721 aagttagggt aggcttcaac aggaaggtca gattgtttaa accaagcggc gactgagatg
     6781 tcctaaatgc acagcgacgg attcgcgcta tttagaaaga gagagcaata tttcaagaat
     6841 gcatgcgtca attttacgca gactatcttt ctagggttaa tctagctgca tcaggatcat
     6901 atcgtcgggt cttttttccg gctcagtcat cgcccaagct ggcgctatct gggcatcggg
     6961 gaggaagaag cccgtgcctt ttcccgcgag gttgaagcgg catggaaaga gtttgccgag
     7021 gatgactgct gctgcattga cgttgagcga aaacgcacgt ttaccatgat gattcgggaa
     7081 ggtgtggcca tgcacgcctt taacggtgaa ctgttcgttc aggccacctg ggataccagt
     7141 tcgtcgcggc ttttccggac acagttccgg atggtcagcc cgaagcgcat cagcaacccg
     7201 aacaataccg gcgacagccg gaactgccgt gccggtgtgc agattaatga cagcggtgcg
     7261 gcgctgggat attacgtcag cgaggacggg tatcctggct ggatgccgca gaaatggaca
     7321 tggatacccc gtgagttacc cggcgggcgc gcttggcgta atcatggtca tagctgtttc
     7381 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7441 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7501 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7561 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7621 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7681 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7741 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7801 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7861 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7921 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7981 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     8041 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     8101 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8161 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     8221 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8281 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8341 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     8401 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8461 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8521 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8581 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8641 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8701 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8761 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8821 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8881 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8941 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     9001 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     9061 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9121 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9181 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9241 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9301 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9361 gggcgacacg gaaatgttga atactcat
//
