LOCUS       sequence-12495-e        6114 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 6114)
  AUTHORS   Held M, Schmitz MH, Fischer B, Walter T, Neumann B, Olma MH, Peter 
            M, Ellenberg J, Gerlich DW
  TITLE     CellCognition: time-resolved phenotype annotation in high-throughput
            live cell imaging.
  JOURNAL   Nat Methods. 2010 Sep . 7(9):747-54.
  PUBMED    20693996
REFERENCE   2  (bases 1 to 6114)
  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: "Nat 
            Methods. 2010 Sep . 7(9):747-54."
COMMENT     Sequence Label: pGALT-EGFP-IRES-puro2b
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(965..993)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1588..1608
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     intron          1723..1952
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1935..1954
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2010..2584,2585..2587,2588..2596)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2177..2194)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2404..2423
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2616..3215
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2616..2635)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3112..3132
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3351..3368)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3357..3581
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3628..3647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3707..3723)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3707..3723)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3720..3742)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3731..3747
                     /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(join(3755..3761,3762..3779,3780..3785))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3800..3821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3938..3955)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4109..4697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4189..4208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4868..5659,5660..5728))
                     /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     5491..5510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5729..5833)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5908..5927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(6099..6114,1..364)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attccgggcg ggtcgcctcc cctcctgtag
      661 cccacaccct tcttaaagcg gcggcgggaa gatgaggctt cgggagccgc tcctgagccg
      721 gagcgccgcg atgccaggcg cgtccctaca gcgggcctgc cgcctgctcg tggccgtctg
      781 cgctctgcac cttggcgtca ccctcgttta ctacctggct ggccgcgacc tgagccgcct
      841 gccccaactg gtcggagtct ccacaccgct gcagggcggg tcgaacagtg ccgccgccat
      901 cgggcagtcc tccggggacc tccggaccgg agggatgagt aaaggagaag aacttttcac
      961 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
     1021 cagtggagag ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac
     1081 tactggaaaa ctacctgttc catggccaac cctggtcacc accctgacct acggcgtgca
     1141 gtgcttctcc cgttaccctg atcatatgaa gcggcacgac ttcttcaaga gcgccatgcc
     1201 tgagggatac gtgcaggaga ggaccatctt cttcaaggac gacgggaact acaagacacg
     1261 tgctgaagtc aagtttgagg gagacaccct cgtcaacagg atcgagctta agggaatcga
     1321 tttcaaggag gacggaaaca tcctcggcca caagttggaa tacaactaca actcccacaa
     1381 cgtatacatc atggccgaca agcaaaagaa cggcatcaaa gccaacttca agacccgcca
     1441 caacatcgaa gacggcggcg tgcaactcgc tgatcattat caacaaaata ctccaattgg
     1501 cgatggccct gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa
     1561 agatcccaac gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat
     1621 tacacatggc atggatgaac tatacaaatg aacgcgggcc gcatagataa ctgatccagt
     1681 gtgctggaat taattcgctg tctgcgaggg ccagctgttg gggtgagtac tccctctcaa
     1741 aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat
     1801 tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca gaaaagacaa
     1861 tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac acttgagtga
     1921 caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcaggt
     1981 cgagcatgca tctagggcgg ccaattccgc ccctctccct cccccccccc taacgttact
     2041 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgtgatt ttccaccata
     2101 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
     2161 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
     2221 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
     2281 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
     2341 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
     2401 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
     2461 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
     2521 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
     2581 taagcttgcc acaacccaca aggagacgac cttccatgac cgagtacaag cccacggtgc
     2641 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     2701 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     2761 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     2821 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     2881 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     2941 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     3001 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     3061 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     3121 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgagtgcccg aaggaccgcg
     3181 cgacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc
     3241 cgaccgaaag gagcgcacga ccccatggct ccgaccgaag ccgacccggg cggccccgcc
     3301 gaccccgcac ccgcccccga ggcccaccga ctctagagct cgctgatcag cctcgactgt
     3361 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3421 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3481 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3541 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3601 cagctggggc tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3661 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3721 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3781 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3841 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3901 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3961 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4021 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4081 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     4321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4921 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4981 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5041 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5101 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5161 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5221 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5281 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5341 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5401 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5461 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5521 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5581 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5641 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5701 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5821 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     5881 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     5941 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6001 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6061 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgac
//
