LOCUS       sequence-60296-e        5486 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 5486)
  AUTHORS   Piatkevich KD, Subach FV, Verkhusha VV
  TITLE     Far-red light photoactivatable near-infrared fluorescent proteins 
            engineered from a bacterial phytochrome.
  JOURNAL   Nat Commun. 2013 Jul 10;4:2153. doi: 10.1038/ncomms3153.
  PUBMED    23842578
REFERENCE   2  (bases 1 to 5486)
  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 
            Commun."; date: "2013-07-10"; volume: "4:2153. doi"; pages: " 
            10.1038/ncomms3153"
COMMENT     Sequence Label: pPAiRFP2-N1
FEATURES             Location/Qualifiers
     source          1..5486
                     /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      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2274..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2311..2330)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2365..2384
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2402..2857)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2539..2560)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2751..2770
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2884..2988
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2990..3347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2990..3181
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3013..3033)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3198..3333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3382..4176
                     /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             3382..4176
                     /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(3436..3455)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4046..4065
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4364..4383)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4408..4455
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4784..5372
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5273..5292
                     /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 atctcgccac catggcatcc acggactatc atgtcgacct gaccaattgc
      661 gaccgcgaac ccattcatat tccaggatat attcaaccgc atggctgcct catcgcctgc
      721 gataacgcca tgcgaatggt gctgcggcat tccgaaaact gcggcgaact gctgggactt
      781 gagggcgatc tcaacggcag gaccgccgag gatgtgctcg gcaaaaggct cgtccatgat
      841 ctccgcaacg cgctcaccgt caccggtaaa accacgcgcc ccgccatgct gcccgcaatg
      901 gaaacaagcg acggccgcag cttcgatatt tcactccatc gctacaaatc caccaccatc
      961 atcgaattcg agccctcgga cagcgacacg cagccgctcg gcacggcgcg caagatggtg
     1021 gaccgcatcc gcgaagccga cagcgtcgaa agcctgatct ccagaaccac acgcctggta
     1081 aaggcgacgc tgggttacga ccgtgtgctg atctaccgct tcgaggaaga cggcgccggc
     1141 aaggtcgtgt cggaggccaa gcagcccgag ctggagagct ttctcgggca atatttcccc
     1201 gccagcgata ttccacagca ggctcgcgca ctttatctca agaacacctt gcgcatcatc
     1261 tccgatgcca gcggcacccc cattccggtc ctgccagccg tcgatgtctc cggtgagccg
     1321 ctcgatctct cttacgcgca tttgcgcaac ttctcgccca tccattgcga atatctccgc
     1381 aatatgggtg ttgccgcctc catgtcgatt tctgtgattg tcgatgacgc cctgtggggc
     1441 ctgatcgtct gccaccattg ttcgccgcgt gtgctgagca tgccagttcg gatcgctgcg
     1501 gcgatgttcg gggaattttt ctccatgttc ctgcaggttc tgaaacagaa gcgccgcctc
     1561 gacaccatca accacgccca tgcggcgctc gaccggtttc tgcggctggc cgcccatcgg
     1621 gccaacctcg aagaactgct ggtcgacagt tttcaggatt ttgcccgcct gataccctgc
     1681 gacggtgtcg ggctgtgggt cggcaacaac tggcacggcc atggcgcaac gccaccgcat
     1741 gatgccattc caaggctggc acgtttcgtc gcttcggttt cggaaggcag ggtctgggca
     1801 acacacgccc tgtcgcaggc cataccggaa gcggaaatat acgccgacac cgcggccgga
     1861 atgcttgcta tcccgatttc gcaggtcaag agcgactatc tcctgttttt ccgcaaggag
     1921 atcgtgcaga acctgaactg ggccggcaat ccggaaaaat cctatgaaac cggcccgatg
     1981 ggcgaccgcc tcacgccgcg taaaagcttc gcaatctgga aagagtccgt tcgcctgcag
     2041 gcacagccct ggtcggaagc cgaccgggaa atcgccgaaa ccgcgagaat cgcgcttgtc
     2101 ggagtggcgt tccaccacag cgagttgatg gcaggcctcg agtacaagta aagcggccgc
     2161 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2221 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2281 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2341 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     2401 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     2461 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     2521 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     2581 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     2641 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     2701 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     2761 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     2821 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     2881 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2941 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3001 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3061 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     3361 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3421 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3481 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3541 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     3601 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3661 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3721 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3781 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3841 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3901 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3961 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4021 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4081 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4141 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4201 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4261 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4321 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     4381 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     4441 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     4501 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     4561 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     4621 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4681 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     4741 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4801 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4861 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4921 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4981 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5041 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5101 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5161 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5221 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5281 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     5341 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     5401 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5461 tggataaccg tattaccgcc atgcat
//
