LOCUS       sequence-50659-e        4718 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 4718)
  AUTHORS   Filonov GS, Verkhusha VV
  TITLE     A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein
            Interactions.
  JOURNAL   Chem Biol. 2013 Jul 23. pii: S1074-5521(13)00242-1. doi: 
            10.1016/j.chembiol.2013.06.009.
  PUBMED    23891149
REFERENCE   2  (bases 1 to 4718)
  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: "Chem Biol. 
            2013 Jul 23. pii: S1074-5521(13)00242-1. doi: 
            10.1016/j.chembiol.2013.06.009."
COMMENT     Sequence Label: pK-GAFm
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    180..301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(217..236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     271..290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(308..763)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(445..466)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     657..676
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        790..894
                     /gene="bla"
                     /label=AmpR promoter
     promoter        896..1253
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        896..1087
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(919..939)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1104..1239
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1288..2082
                     /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             1288..2082
                     /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(1342..1361)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1952..1971
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2270..2289)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2314..2361
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2690..3278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3179..3198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3453..3756
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3757..3960
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3911..3931
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3992..4001
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(4718,1..56)
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccaccg
       61 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
      121 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
      181 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      241 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      301 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
      361 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
      421 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
      481 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
      541 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
      601 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
      661 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
      721 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
      781 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      841 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
      901 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
      961 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     1021 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     1081 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     1141 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     1201 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     1261 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     1321 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     1381 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     1441 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     1501 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     1561 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     1621 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     1681 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     1741 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     1801 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     1861 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     1921 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     1981 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     2041 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     2101 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     2161 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     2221 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     2281 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     2341 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     2401 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     2461 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     2521 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2581 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2641 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2701 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2761 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2821 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2881 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2941 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3001 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3061 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3121 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3181 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3241 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3301 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3361 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg
     3421 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     3481 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     3541 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     3601 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     3661 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     3721 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     3781 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     3841 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     3901 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     3961 ggtttagtga accgtcagat ccgctagcgc cgccaccatg ggcggcagca aggtgtctgc
     4021 acttaaagaa aaggtttccg cactcaagga gaaagtttca gctttgaaag agaaggtcag
     4081 tgcccttaag gaaaaagtga gcgcgctgaa agaaggcggc agccctcccc agcgggacgt
     4141 cgccgagccg caggcgttct tccgccgcac caacagcgcc atccgccgcc tgcaggccgc
     4201 cgaaaccttg gaaagcgcct gcgccgccgc ggcgcaagag gtgcggaaga ttaccggcta
     4261 cgatcgggtg atgatctatc gcttcgcctc cgacttcagc ggcgaagtga tcgcagagga
     4321 tcggtgcgcc gaggtcgagt caaaactagg cctgcactat cctgcctcaa ccgtgccggc
     4381 gcaggcccgt cggctctata ccatcaaccc ggtacggatc attcccgata tcaattatcg
     4441 gccggtgccg gtcaccccat acctcaatcc ggtcaccggg cggccgattg atcttagctt
     4501 cgccatcctg cgcagcgtct cgcccgtcca tctggaattc atgcgcaaca taggcatgca
     4561 cggcacgatg tcgatctcga ttttgcgcgg cgagcgactg tggggattga tcgtttgcca
     4621 tcaccgaacg ccgtactacg tcgatctcga tggccgccaa gcctgcgagc tagtcgccca
     4681 ggttctggcc cggcagatcg gcgtgatgga agagtaaa
//
