LOCUS       sequence-364497-        6096 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 6096)
  AUTHORS   Howarth M, Takao K, Hayashi Y, Ting AY
  TITLE     Targeting quantum dots to surface proteins in living cells with 
            biotin ligase.
  JOURNAL   Proc Natl Acad Sci U S A. 2005 May 24. 102(21):7583-8.
  PUBMED    15897449
REFERENCE   2  (bases 1 to 6096)
  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: "Proc Natl 
            Acad Sci U S A. 2005 May 24. 102(21):7583-8."
COMMENT     Sequence Label: pDisplay-AP-CFP-TM
FEATURES             Location/Qualifiers
     source          1..6096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(81..101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     328..347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             446..1240
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             446..1240
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(500..519)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1110..1129
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1428..1447)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1472..1519
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1848..2436
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2337..2356
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2568..2792
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2781..2798
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2820..2966)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(2970..2999)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(join(3021..3731,3732..3734,3735..3737))
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3063..3084)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3412..3431
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3671..3692
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3747..3791)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(3819..3845)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     sig_peptide     complement(3849..3908)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     complement(3988..4007)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3989..4007)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4052..4255)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4081..4101)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4256..4635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4647..5102)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4784..4805)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4996..5015
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5129..5233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5234..5302,5303..6094)
                     /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(5452..5471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtcagaccaa gtttactcat atatacttta gattgattta ttgaaaaagg aagagtcctg
       61 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
      121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
      181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
      241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
      301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccc ggcctctgag
      361 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
      421 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      481 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      541 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      601 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
      661 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      721 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      781 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      841 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      901 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
      961 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1021 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1081 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1141 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1201 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1261 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     1321 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1381 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     1441 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     1501 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     1561 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     1621 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     1681 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1741 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1801 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1861 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1921 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1981 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2041 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2101 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2161 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2221 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2281 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2341 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2401 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     2461 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     2521 tctgtggata accgtattac cgccactggt tctttccgcc tcagaagcca tagagcccac
     2581 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     2641 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     2701 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     2761 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatctcgagc ggccgcctaa
     2821 cgtggcttct tctgccaaag catgatgagg atgataaggg agatgatggt gagcaccacc
     2881 agggccagga tggctgagat caccaccacc ttaaagggca aggagtgtgg caccacgatg
     2941 acctcctgcg tgtcctggcc cacagcattc agatcctctt ctgagatgag tttttgttcg
     3001 tcgacctgca gccgcggccg cttgtatagc tcgtccatgc cgagagtgat cccggcggcg
     3061 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3121 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3181 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     3241 aagtgggcct tgatgccgtt cttctgcttg tcggcggtga tatagacgtt gtggctgatg
     3301 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3361 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3421 ttgccgtcgt ccttgaagaa gatggtacgc tcctggacgt agccttcggg catggcggac
     3481 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccccag
     3541 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     3601 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacctg
     3661 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     3721 tcgcccttgc tcaccatcgg cgcgccctcg tgccactcga tcttctgggc ctcgaagatg
     3781 tcgttcaggc cgccgccaga tctggccggc tgggccccag cataatctgg aacatcatat
     3841 ggatagtcac cagtggaacc tggaacccag agcagcagta cccatagcag gagtgtgtct
     3901 gtctccatgg tggatatccc caagccgaat tccagcacac tggcggccgt tactagtgga
     3961 tccgagctcg gtaccaagct tgggtctccc tatagtgagt cgtattaatt tcgataagcc
     4021 agtaagcagt gggttctcta gttagccaga gagctctgct tatatagacc tcccaccgta
     4081 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     4141 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4201 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4261 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4321 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4381 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4441 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     4501 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     4561 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     4621 tcaataatca atgtcaacgc gcgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     4681 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     4741 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     4801 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     4861 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     4921 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     4981 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     5041 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     5101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5221 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5281 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5341 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5401 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5461 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5521 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5581 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5641 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5701 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5761 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5821 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5881 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5941 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6001 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6061 tgagataggt gcctcactga ttaagcattg gtaact
//
