LOCUS       sequence-329824-        6056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EBFP2(1-10) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6056)
  AUTHORS   Tamura R, Jiang F, Xie J, Kamiyama D
  TITLE     Multiplexed labeling of cellular proteins with split fluorescent 
            protein tags.
  JOURNAL   Commun Biol. 2021 Feb 26;4(1):257. doi: 10.1038/s42003-021-01780-4.
  PUBMED    33637968
REFERENCE   2  (bases 1 to 6056)
  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: "Commun 
            Biol."; date: "2021-02-26"; volume: "4(1):257. doi"; pages: " 
            10.1038/s42003-021-01780-4"
COMMENT     Sequence Label: pcDNA3.1-EBFP2(1-10)
FEATURES             Location/Qualifiers
     source          1..6056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(740..761)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1413..1430)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1419..1643
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1689..2117
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1776..1795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1986..2007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2126..2146)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2131..2460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2311..2446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2373..2392
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2527..3321
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2581..2600)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3191..3210
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3532..3551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3586..3605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3665..3681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3665..3681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3678..3700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3689..3705
                     /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(3713..3719,3720..3737,3738..3743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3758..3779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3896..3913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4067..4655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4147..4166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4826..5617,5618..5686))
                     /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     5449..5468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5687..5791)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5866..5885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccgg atccatggtg agcaagggcg aggagctgtt
      721 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      781 cgtgaggggc gagggcgagg gcgatgccac catcggcaag ctgaccctga agttcatctg
      841 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt
      901 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      961 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca aatacaagac
     1021 ccgcgccgtg gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcgt
     1081 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca
     1141 caacatctat atcatggccg tcaagcagaa gaacggcatc aaggtgaact tcaccgtgcg
     1201 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1261 cggcgacggc cccgtgctgc tgcccgacag ccactacctg agcacccaga ccgtgctgag
     1321 caaagacccc aacgagaagt aagaattctg cagatatcca gcacagtggc ggccgctcga
     1381 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     1441 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1501 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1561 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1621 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     1681 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1741 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1801 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1861 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1921 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1981 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2041 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2101 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     2161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     2221 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2281 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     2341 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     2401 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     2461 aagctcccgg gagcttatat atccattttc ggatctgatc aagagacagg atgaggatcg
     2521 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2581 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2641 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2701 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2761 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2821 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2881 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2941 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3001 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3061 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3121 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3181 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3241 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3301 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3361 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3421 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3481 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3541 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3601 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3661 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3721 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3781 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3841 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3901 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3961 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4021 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4081 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4141 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4201 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4261 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4321 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4381 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4441 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4501 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4561 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4621 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4681 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4741 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4801 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4861 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4921 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4981 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5041 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5101 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5161 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5221 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5281 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5341 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5401 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5461 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5521 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5581 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5641 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5701 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5761 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5821 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     5881 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5941 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6001 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
