LOCUS       sequence-35862-e        5622 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 5622)
  AUTHORS   Clark KJ, Balciunas D, Pogoda HM, Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielsen AL, 
            Patowary A, Scaria V, Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC
  TITLE     In vivo protein trapping produces a functional expression codex of 
            the vertebrate proteome.
  JOURNAL   Nat Methods. 2011 Jun;8(6):506-15. Epub 2011 May 8.
  PUBMED    21552255
REFERENCE   2  (bases 1 to 5622)
  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."; date: "2011-06"; volume: "8(6)"; pages: "506-15. Epub 
            2011 May 8"
COMMENT     Sequence Label: pGBT-R16
FEATURES             Location/Qualifiers
     source          1..5622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(2..13,14..176)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    221..254
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             581..1255
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     CDS             581..1255
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR
                     LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST
                     GA"
     primer_bind     complement(708..726)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     976..995
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1170..1193
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    2366..2399
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(2484..2671,2672..2683)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(2695..2714)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2696..2714)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2721..2738)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2721..2737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2730..2752)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2878..3333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2965..2984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3175..3196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3411..3515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3516..3584,3585..4376)
                     /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(3734..3753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4547..5135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5036..5055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5289..5306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5423..5444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5459..5464,5465..5482,5483..5489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5497..5513
                     /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     5502..5524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5521..5537
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5521..5537
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5555..5573
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5555..5572
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gcagaggtgt aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa
       61 aattttactc aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa
      121 aagtactcca ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa
      181 aactagagat tcttgtttaa gctgtaggga taacagggta ataacttcgt ataatgtatg
      241 ctatacgaag ttatcagcat gcaaaccgtt ctctgaagcc tctcgtctgt cccgtttcct
      301 cattttaagt tctcctgttc attggttcac ttcctccagg ctttgtgtcc tctgagctcc
      361 tgagtttctc ctcttctcag caggttatct aaactttgcc tgcctgtttt gcagttcctt
      421 ctgcactttt tgttccttta ctctggattt ccgactaacc cctgcatgga agtgtggatt
      481 gtgtgctgca gacacttgaa catccagtta acgtctccat cctctctttg cagatcatgt
      541 tggagacctt caacaccccc gcccccgggt accggaaggt gcctcctccg aggacgtcat
      601 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
      661 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      721 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg
      781 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
      841 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      901 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      961 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     1021 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa
     1081 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca
     1141 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta
     1201 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaagaaat
     1261 cgatacggac tgttaccact tcacgccgac tcaaactgcg cagagaaact tcaaacgaca
     1321 acattggcat ggcttttgtt atttttggcg cttgactcag gatctaaaaa ctggaacggt
     1381 gaaggtgacg gcaatgtttt ttggcaaata agcatccccg aagttctaca atgcatctga
     1441 ggactccatg ttttgttttc tttagtcatt ccaaatgttt gttaaatgca ttgttccgaa
     1501 acttatttgc ctctatgaag gctgcccagt aattgggagc atacttaaca ttgtagtatt
     1561 gtatgtaaat tatgtaacaa aacaatgtct gggtttttgt acttttcagc cttaaaatct
     1621 tgggtttatt tttttttctt tgttccaaaa aaacgaagct ttaccattca agatgtaaag
     1681 gtttccattc cccctgggca tattgtaaaa gctgtgtgga acgtggcggt gccagacatt
     1741 tggtggggcc aacctgtaca ctgactaatt caattccaat aaaagtgcac atgtaagaca
     1801 tcctacccct gtctcatttc ctgtcttggc tctgagtggc ttcaataaaa atggcttttg
     1861 tgaagttgca ccatgttaat tacatttcag cactttcaca ccaggttttt gactaatttg
     1921 aatttatcga agtttgacct tttttttttt ttttccttca agctaatttg aggttatcaa
     1981 accatttcat tggaggaaac ttaatgttta aaagcataaa ttcgaaacat gattcattta
     2041 tttaactgct taatggtgaa acaaacttca agtaatacac caagtgttcc ctcatatgaa
     2101 ggtagctaac tgcaaacggc cttaactttc catcttaagt tgagctacat aaaggcttta
     2161 acaggaagga agagccagtt taaaagtcaa accaccatga ctgatataaa ttcttgtcca
     2221 tataaccttg cgtttcacac aatatagcta tctactgcta aataaattat cgcgattcct
     2281 attctctaga aagtatagga acttcctagg agcgctccgg agatcttcct attctctaga
     2341 aagtatagga acttcttaat taacgataac ttcgtataat gtatgctata cgaagttata
     2401 ttaccctgtt atccctacca tgaaattaaa ctgggcatca gcgcaattca attggtttgg
     2461 taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa
     2521 ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt
     2581 aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa
     2641 gtacagtaat caagtaaaat tactcaagta ctttacacct ctgggcccaa ttcgccctat
     2701 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2761 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2821 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     2881 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2941 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3001 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3061 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3121 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3181 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3241 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3301 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3361 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3481 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3541 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3601 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3661 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3721 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3781 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3841 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3901 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3961 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4021 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4081 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4141 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4201 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4261 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4321 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4381 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4441 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4501 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4561 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4621 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5161 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5221 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5281 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5341 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5401 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5461 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5521 caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat agaatactca
     5581 agctatgcat ccaacgcgtt gggagctctc ccatatggtc ga
//
