LOCUS       sequence-8245-ex        4031 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 4031)
  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 4031)
  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#715
FEATURES             Location/Qualifiers
     source          1..4031
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     477..497
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1471..1493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1593..1612
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1690..1708)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1776..1880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1881..1949,1950..2741)
                     /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(2099..2118)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2912..3500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3401..3420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3654..3671
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3788..3809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3824..3829,3830..3847,3848..3854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3862..3878
                     /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     3867..3889
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3886..3902
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3886..3902
                     /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 aaatggctag cgcacagctg gagaagaaac tgcaggctct ggagaagaaa
      181 ctggcacagc tggagtggaa aaaccaggca ctggagaaga aactggcaca gggtggaagc
      241 ggtaagaatg ggatcaaagc ggtaagttta aacatgattt tactaactaa ctaatctgat
      301 ttaaattttc agaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
      361 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
      421 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
      481 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaatag
      541 cattcgtaga attccaactg agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa
      601 aataataggg gccgctgtca tcagagtaag tttaaactga gttctactaa ctaacgagta
      661 atatttaaat tttcagcatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
      721 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
      781 aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
      841 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
      901 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
      961 gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg cacacaaaat
     1021 accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg
     1081 gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt
     1141 ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg ggggtttccc
     1201 ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
     1261 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
     1321 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
     1381 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
     1441 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1501 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1561 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     1621 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     1681 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     1741 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1801 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1861 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1921 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1981 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     2041 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     2101 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     2161 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     2221 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     2281 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2341 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2401 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2461 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2521 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2581 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2641 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2701 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2761 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2821 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2881 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2941 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3001 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     3061 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     3121 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     3181 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     3241 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3301 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3361 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3421 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3481 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3541 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3601 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3661 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3721 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     3781 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     3841 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     3901 accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact attctcattt
     3961 aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg ctaacaactt
     4021 ggaaatgaaa t
//
