LOCUS       sequence-8238-ex        4104 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 4104)
  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 4104)
  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#707
FEATURES             Location/Qualifiers
     source          1..4104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..98)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             583..594
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(1544..1566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1666..1685
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1763..1781)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1849..1953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1954..2022,2023..2814)
                     /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(2172..2191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2985..3573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3474..3493
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3727..3744
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3861..3882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3897..3902,3903..3920,3921..3927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3935..3951
                     /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     3940..3962
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3959..3975
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3959..3975
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccatgg ctagcaaagg agaagaactc
       61 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaacgg ccacaagttc
      121 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct gaagttcatc
      181 tgcactactg gcaaactgcc tgttccatgg ccaacactag tcactactct gtgctatggt
      241 gttcaatgct tttcaagata cccggatcat atgaaacggc atgacttttt caagagtgcc
      301 atgcccgaag gttatgtaca ggaaaggacc atcttcttca aagatgacgg caactacaag
      361 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
      421 attgacttca aggaagatgg caacattctg ggacacaaat tggaatacaa ctataactca
      481 cacaacgttc ccatcatggc agacaaacaa ggtggctctg gctctggcgc tcagcttaag
      541 aaagagctgc aggcaaacaa gaaagagctg gctcagctga agtgggaact gcaggcactg
      601 aagaaagaac tggctcagtg agaattccaa ctgagcgccg gtcgctacca ttaccaactt
      661 gtctggtgtc aaaaataata ggggccgctg tcatcagagt aagtttaaac tgagttctac
      721 taactaacga gtaatattta aattttcagc atctcgcgcc cgtgcctctg acttctaagt
      781 ccaattactc ttcaacatcc ctacatgctc tttctccctg tgctcccacc ccctattttt
      841 gttattatca aaaaaacttc ttcttaattt ctttgttttt tagcttcttt taagtcacct
      901 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
      961 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     1021 gtacacttct tatgtttttt ttacttctga taaatttttt ttgaaacatc atagaaaaaa
     1081 ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc
     1141 ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt
     1201 atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt
     1261 ttgggggttt cccctattgt ttgtcaagag tttcgaggac ggcgtttttc ttgctaaaat
     1321 cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca
     1381 gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc
     1441 ttaaatgaca gaatacattc ccaatatacc aaacataact gtttcctact agtcggccgt
     1501 acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1741 accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1801 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1861 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1921 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1981 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2041 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2101 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2161 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2221 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2281 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2341 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2401 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2461 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2521 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2581 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2641 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2701 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2761 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2821 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2881 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2941 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     3001 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     3061 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     3121 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     3181 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     3241 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     3301 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     3361 tacctacagc gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     3421 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     3481 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     3541 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     3601 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     3661 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     3721 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     3781 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     3841 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     3901 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     3961 ggaaacagct atgaccatga ttacgccaag ctgtaagttt aaacatgatc ttactaacta
     4021 actattctca tttaaatttt cagagcttaa aaatggctga aatcactcac aacgatggat
     4081 acgctaacaa cttggaaatg aaat
//
