LOCUS       sequence-79080-e        7508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  scaffold for PI4KA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7508)
  AUTHORS   Hammond GR, Machner MP, Balla T
  TITLE     A novel probe for phosphatidylinositol 4-phosphate reveals multiple 
            pools beyond the Golgi.
  JOURNAL   J Cell Biol. 2014 Apr 7.
  PUBMED    24711504
REFERENCE   2  (bases 1 to 7508)
  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: "J Cell 
            Biol. 2014 Apr 7."
COMMENT     Sequence Label: iRFP-TTC7B
FEATURES             Location/Qualifiers
     source          1..7508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     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"
     regulatory      3217..3226
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3223..4173
                     /codon_start=1
                     /product="phytochrome-based near-infrared fluorescent 
                     protein, originally termed iRFP (Filonov et al., 2011)"
                     /label=iRFP713
                     /note="name was changed to iRFP713 (Shcherbakova and 
                     Verkhusha, 2013)"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGEVIAEDRCAEVESKLGLHYPASTVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPVHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    4296..4417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4333..4352)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4387..4406
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4424..4879)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4561..4582)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4773..4792
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4906..5010
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5012..5369
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5012..5203
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5035..5055)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5220..5355
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5404..6198
                     /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             5404..6198
                     /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(5458..5477)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6068..6087
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6386..6405)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6430..6477
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6806..7394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7295..7314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag 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 caagcttacc atggcgacca agaaggcagg ctcgcggctg
      661 gagacggaga tcgagcgctg ccgctccgag tgccagtggg agcggatccc tgagctcgtc
      721 aagcagctgt cggccaagct catcgccaac gatgacatgg cagagcttct cctcggggag
      781 tcgaagctgg agcagtacct gaaggaacac cccctgaggc agggggccag tccccgaggc
      841 cccaagcccc agctgactga ggtccgcaag catctgaccg ccgccctgga ccgagggaac
      901 cttaagtcag aattcctaca agaatccaat ctgatcatgg ccaagttgaa ttatgtggaa
      961 ggtgattata aagaagctct gaacatttac gcccgggtgg gcctggacga tctgccactg
     1021 acagctgtcc cgccctacag gctgcgggtg atcgcagaag cctacgctac caaaggactt
     1081 tgtttggaga agctgcctat ttcttcttct accagtaatc tccatgtgga ccgggaacag
     1141 gatgtcatca cctgttatga gaaagcaggg gacatcgcac tcctgtatct ccaagagata
     1201 gaaagggtaa tactttctaa tattcaaaac agaagcccta agcctggccc tgctccccac
     1261 gatcaagaac taggtttttt cctagaaaca ggacttcaga gagcccatgt cctctatttc
     1321 aaaaatggga acttgacaag aggagtcgga agatttagag agcttctcag agcagttgaa
     1381 acaagaacga ctcaaaacct gcgaatgaca atagccaggc agctggcaga gatcttgttg
     1441 cggggtatgt gtgagcagag ctactggaac cctctggagg atccaccgtg ccagtcacct
     1501 ctggacgatc ctctccgcaa aggagcaaac acaaaaacct acactctcac tcggagagcc
     1561 cgtgtctact caggagagaa cattttttgt cctcaagaaa atacggaaga agccctgttg
     1621 ttattgctga ttagtgaatc aatggccaac cgggacgctg tgctgagcag gatacctgaa
     1681 cacaagagtg accgcctcat cagtctgcag agtgcatctg tggtctatga cttactcacc
     1741 attgctcttg gaagaagagg ccagtatgag atgctgtcag agtgcctaga aagagccatg
     1801 aagtttgcct ttgaggaatt ccacctgtgg taccagtttg ctctgtccct gatggctgct
     1861 ggaaaatctg cccgtgccgt gaaggtgctg aaagagtgta tccgcctgaa gccagacgat
     1921 gccaccatcc ctctcctcgc tgccaagctc tgcatgggct ccctgcactg gttggaagag
     1981 gctgaaaagt ttgccaaaac tgtcgttgat gtgggagaga aaacgtcaga gttcaaggcc
     2041 aaaggctact tagctctggg gctcacgtac agtctgcagg ccactgacgc ttctttgcga
     2101 gggatgcagg aggtcctaca gagaaaggcg cttcttgcat ttcagagggc ccacagcctg
     2161 tcacccacag atcaccaagc agctttctac ctggctctgc agcttgccat ctccagacag
     2221 atcccagagg ctctggggta tgtccgccaa gctcttcagc ttcaaggtga cgatgccaac
     2281 tccctgcacc tccttgccct cctgctgtca gcacagaagc attaccatga cgctctgaac
     2341 atcatcgaca tggccctgag tgaataccca gaaaatttca tactactgtt ttccaaagtg
     2401 aagttgcagt cactctgccg aggcccggac gaggcactgc tgacttgtaa gcacatgctg
     2461 cagatatgga aatcctgcta caacctcacc aaccccagtg attctggacg tgggagcagc
     2521 ctcttagata gaaccattgc tgacagacga cagcttaata caattacttt gccagacttc
     2581 agcgatcccg agacaggctc cgtccatgcc acatcggtag cagcctcaag agtggagcag
     2641 gcactgtcgg aagtggcttc gtctctgcag agcagtgccc ctaagcaggg cccgctgcac
     2701 ccctggatga cgctggcaca gatctggctc catgcagctg aagtctatat cggcatcggg
     2761 aagcctgcag aagccacagc ctgtacccaa gaagctgcca acctcttccc aatgtcccac
     2821 aatgtcctct acatgcgcgg ccagattgct gagctccggg gaagcatgga cgaggcgcgg
     2881 cggtggtatg aagaggcctt agccatcagc cccacccacg tgaagagcat gcagcgactg
     2941 gccctgatcc ttcaccagct aggccgctac agtctggcgg agaagatcct ccgggacgcg
     3001 gtgcaggtga actcgacagc ccacgaggtc tggaacgggc tgggcgaggt cctccaagct
     3061 cagggcaacg atgcggcggc tacggagtgc ttcctgacag ccttggagct ggaggccagc
     3121 agccccgccg tgcccttcac catcatcccc cgcgtgctcg gtggtagtgc tggtggtagt
     3181 gcgtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggcgga aggatccgtc
     3241 gccaggcagc ctgacctctt gacctgcgac gatgagccga tccatatccc cggtgccatc
     3301 caaccgcatg gactgctgct cgccctcgcc gccgacatga cgatcgttgc cggcagcgac
     3361 aaccttcccg aactcaccgg actggcgatc ggcgccctga tcggccgctc tgcggccgat
     3421 gtcttcgact cggagacgca caaccgtctg acgatcgcct tggccgagcc cggggcggcc
     3481 gtcggagcac cgatcactgt cggcttcacg atgcgaaagg acgcaggctt catcggctcc
     3541 tggcatcgcc atgatcagct catcttcctc gagctcgagc ctccccagcg ggacgtcgcc
     3601 gagccgcagg cgttcttccg ccgcaccaac agcgccatcc gccgcctgca ggccgccgaa
     3661 accttggaaa gcgcctgcgc cgccgcggcg caagaggtgc ggaagattac cggcttcgat
     3721 cgggtgatga tctatcgctt cgcctccgac ttcagcggcg aagtgatcgc agaggatcgg
     3781 tgcgccgagg tcgagtcaaa actaggcctg cactatcctg cctcaaccgt gccggcgcag
     3841 gcccgtcggc tctataccat caacccggta cggatcattc ccgatatcaa ttatcggccg
     3901 gtgccggtca ccccagacct caatccggtc accgggcggc cgattgatct tagcttcgcc
     3961 atcctgcgca gcgtctcgcc cgtccatctg gaattcatgc gcaacatagg catgcacggc
     4021 acgatgtcga tctcgatttt gcgcggcgag cgactgtggg gattgatcgt ttgccatcac
     4081 cgaacgccgt actacgtcga tctcgatggc cgccaagcct gcgagctagt cgcccaggtt
     4141 ctggcctggc agatcggcgt gatggaagag tgaagcggcc gcgactctag atcataatca
     4201 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4261 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4321 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4381 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     4441 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4501 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4561 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4621 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4681 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4741 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4801 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4861 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4921 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4981 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5041 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5101 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5161 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5221 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5281 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5341 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5401 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5461 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5521 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5581 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5641 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5701 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5761 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5821 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5881 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5941 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6001 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6061 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6121 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6181 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6241 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6301 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6361 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6421 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6481 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6541 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6601 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6661 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6721 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6781 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6841 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6901 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6961 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7021 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7081 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7141 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7201 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7261 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7321 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7381 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7441 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7501 ccatgcat
//
