LOCUS       Exported               10256 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 10256)
  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..10256
                     /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..5253
                     /label=COX7A2
     misc_feature    4187..5186
                     /label=Promoter 1000bp upstream
     misc_feature    5187..5253
                     /label=5' UTR
     misc_feature    5254..5262
                     /label=Exon_1
     misc_feature    5263..6139
                     /label=COX5B intron
     misc_feature    6140..6145
                     /label=same as cox5b exon2
     CDS             6146..6373
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="LKGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDI
                     TSHNEDYTIVEQYERAEGRHSTGGMDELYK"
     misc_feature    6374..6516
                     /label=3' UTR
     misc_feature    6517..7516
                     /label=1000 bp downstream
     misc_feature    7710..7744
                     /label=3'ITR
     primer_bind     complement(8235..8251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8259..8275
                     /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(8283..8313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8328..8349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8637..9225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9396..10256)
                     /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 aagatgagcg aggtactcgg gtctccgggc
     5281 gtgccaggga ccagagtgtt gccctcccag ggtggtccca gggcggcaaa gcggcgcggc
     5341 tcgtgcagct tctcgaggtc ccagtggccg ctttacggtc cccagtgcct caggctctgc
     5401 aggcatctcc ctgtaattct ggaccgctgc tcctgccgct ccccgaactc actccgctgc
     5461 gaaagtatcc taaacggagg tgccgggtga ccttgggagg gaccggggct gccaccggga
     5521 tggggagggg tccggcctcc cttcaaacct gcgcccacct caagcagagt gggttctaca
     5581 tgcttttaga caaatgtcga caaatttgcc tcggtggttg gagaaagaaa agctcatagg
     5641 ccgggcgcgg tggctcacaa ctgtaatccc agcactttgg gaggccgagg cggacagatc
     5701 ccctgaggtc aggagctcaa gaccagcctg gccaacatgg taaaaccccg tttctactaa
     5761 aaatacaaaa attagccggg cgtggtggcg cgcgcctgta gtcccagcta ctcgggaggc
     5821 tgaggcagga gaatcgctta aacccgggag gcggaggttc ccgtgagcca acatcgcgcc
     5881 attgcactcc agcctgggca acaagagcaa aactccgtct caaaaaagaa aaaaaaagct
     5941 cccccgagtg ctgccgcttg tgtggatggg tacttggtgg ttcttagggg accatggata
     6001 tgagtagcct ttaggagctt gtgagcccgc taaaacttat acagaagttt cggggcacca
     6061 ttttccttga tcatttctgt ttgtagtttt tctatcagtc atttcagtca gcgtcataat
     6121 tcacgttatc ttcctttagg tggtgctgaa gggcgagatc aagcagaggc tgaagctgaa
     6181 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     6241 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     6301 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     6361 gctgtacaag taacttcagt catcccagca atcgcttggt tcagtttcat tcagctctct
     6421 atggaccagt aatctgataa ataaccgagc tcttctttgg ggatcaatat ttattgactt
     6481 gtagtaactg ccaccaataa agcagtcttt accatgcttt gtctgatttt atcactttat
     6541 gccaataatt tcaacttttg gtcagccttg taaatcttag gtaaggatca aagaaatgat
     6601 tcttttgcca ctaaacttgt gaaagcaaca ttattgtgat gttattttag tgaaatttta
     6661 cttctaacag ctcaaaatta gtctcaactt ctgtctcaac ttgccgtttg aatgcatatt
     6721 gctttttccc cctacccact ccagtatatc ttggaatatg ctagaacccg ggggtagtgg
     6781 aagcagatta tctttggctc ctggggtcta tatttctttt tttttttttt tttctcagct
     6841 atgtgagagt atgcttaaat attggatata tcccacagct tagaattagg ctttagtaga
     6901 atggctttgt aagatatttc agtatttgtt tttttctaca ctccgagtgg gttcaggtag
     6961 gtttctctca tcaggaacca accaaagcag gacagttcct gggctcccca gctatcatct
     7021 aatggaccca caactgttta acctaaacag tagttaggtg cttaataaat ggtagccata
     7081 ataggctgct cataatagga atgtttcaaa aaggtctatt tagggaacac aaaatccaaa
     7141 tagatacaga gagtaagaat caaaaagaat tacgatgctg ctctaggtag tttgcttcag
     7201 ggtttgcaga acccttcctg ctttgagaga actatgcatg aagctggaaa tttcatagac
     7261 ttcaagtttg ggactccctt ttcctgggtc ctgttgtgca tccatgtgga gagtagcatc
     7321 ttgtgaacca cttcaggact ttccttctat atatatcgag attggtaatt cgtcaagaga
     7381 tactgagtgc ctactgtgtc ctaactggtt ttaggtacct aggaatcact ggtgaacaaa
     7441 agcagattct tcccctcata tgttatagaa aggggtggta ggaaagagag tttaaaaata
     7501 aattgcacag taggttaaaa gataagttct atggaaaaag accaaagggg agaactcaca
     7561 tgggtgtggt tggttggttg cagtattaaa gttagggtag gcttcaacag gaaggtcaga
     7621 ttgtttaaac caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt
     7681 tagaaagaga gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct
     7741 agggttaatc tagctgcatc aggatcatat cgtcgggtct tttttccggc tcagtcatcg
     7801 cccaagctgg cgctatctgg gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt
     7861 tgaagcggca tggaaagagt ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa
     7921 acgcacgttt accatgatga ttcgggaagg tgtggccatg cacgccttta acggtgaact
     7981 gttcgttcag gccacctggg ataccagttc gtcgcggctt ttccggacac agttccggat
     8041 ggtcagcccg aagcgcatca gcaacccgaa caataccggc gacagccgga actgccgtgc
     8101 cggtgtgcag attaatgaca gcggtgcggc gctgggatat tacgtcagcg aggacgggta
     8161 tcctggctgg atgccgcaga aatggacatg gataccccgt gagttacccg gcgggcgcgc
     8221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     8281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     8341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     8461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     8521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     8581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     8641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     8701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     8941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     9001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     9061 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     9121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     9181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     9241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     9301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     9361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     9421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     9481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     9541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     9601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     9661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     9721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     9781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     9841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     9901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     9961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
    10021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
    10081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
    10141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
    10201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcat
//
