LOCUS       sequence-8244-ex        4307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4307)
  AUTHORS   Zhang S, Ma C, Chalfie M
  TITLE     Combinatorial marking of cells and organelles with reconstituted 
            fluorescent proteins.
  JOURNAL   Cell. 2004 Oct 1. 119(1):137-44.
  PUBMED    15454087
REFERENCE   2  (bases 1 to 4307)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2004 
            Oct 1. 119(1):137-44."
COMMENT     Sequence Label: TU#714
FEATURES             Location/Qualifiers
     source          1..4307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(163..191)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             778..789
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1747..1769)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1869..1888
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1966..1984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2052..2156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2157..2225,2226..3017)
                     /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"
     primer_bind     complement(2375..2394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3188..3776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3677..3696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3930..3947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4064..4085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4100..4105,4106..4123,4124..4130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4138..4154
                     /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)."
     primer_bind     4143..4165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4162..4178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4162..4178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttgagggta
      121 ccggtagaaa aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt
      181 gaattagatg gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat
      241 gcaacatacg gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca
      301 tgggtaagtt taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca
      361 cttgtcacta ctttctcatg gggcgttcaa tgcttctcga gatacccaga tcatatgaaa
      421 cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
      481 ttcaaagatg acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca
      541 tacatattta aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga
      601 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
      661 tacaactata tttcacacaa cgtatacatc actgccgaca aacaaggtgg ctctggctct
      721 ggcgctcagc ttaagaaaga gctgcaggca aacaagaaag agctggctca gctgaagtgg
      781 gaactgcagg cactgaagaa agaactggct cagtagcatt cgtagaattc caactgagcg
      841 ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata ataggggccg ctgtcatcag
      901 agtaagttta aactgagttc tactaactaa cgagtaatat ttaaattttc agcatctcgc
      961 gcccgtgcct ctgacttcta agtccaatta ctcttcaaca tccctacatg ctctttctcc
     1021 ctgtgctccc accccctatt tttgttatta tcaaaaaaac ttcttcttaa tttctttgtt
     1081 ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta
     1141 attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc
     1201 ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttttacttc tgataaattt
     1261 tttttgaaac atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag
     1321 tttatgaccg caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc
     1381 tcatcgtgaa aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa
     1441 attaattttc ctgcttttgc tttttggggg tttcccctat tgtttgtcaa gagtttcgag
     1501 gacggcgttt ttcttgctaa aatcacaagt attgatgagc acgatgcaag aaagatcgga
     1561 agaaggtttg ggtttgaggc tcagtggaag gtgagtagaa gttgataatt tgaaagtgga
     1621 gtagtgtcta tggggttttt gccttaaatg acagaataca ttcccaatat accaaacata
     1681 actgtttcct actagtcggc cgtacgggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     1741 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1801 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     1861 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     1921 gcacagatgc gtaaggagaa aataccgcat caggcggcct taagggcctc gtgatacgcc
     1981 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     2041 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2101 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2161 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2221 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2281 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2341 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     2401 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     2461 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2521 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2581 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2641 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2701 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2761 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2821 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2881 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2941 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3001 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3061 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3121 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3181 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3241 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3301 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3361 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3421 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3481 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3541 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     3601 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3661 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3721 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3781 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3841 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3901 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3961 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4021 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     4081 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     4141 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctgtaag
     4201 tttaaacatg atcttactaa ctaactattc tcatttaaat tttcagagct taaaaatggc
     4261 tgaaatcact cacaacgatg gatacgctaa caacttggaa atgaaat
//
