LOCUS       sequence-63499-e        4948 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 4948)
  AUTHORS   Shcherbakova DM, Verkhusha VV
  TITLE     Near-infrared fluorescent proteins for multicolor in vivo imaging.
  JOURNAL   Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521.
  PUBMED    23770755
REFERENCE   2  (bases 1 to 4948)
  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 
            Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521."
COMMENT     Sequence Label: piRFP702-N1
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      7..16
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             13..948
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 702 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP702
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGKVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPIHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     polyA_signal    1070..1191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1107..1126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1161..1180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1198..1653)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1335..1356)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1547..1566
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1680..1784
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1786..2143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1786..1977
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1809..1829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1994..2129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2178..2972
                     /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             2178..2972
                     /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(2232..2251)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2842..2861
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3160..3179)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3204..3251
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3580..4168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4069..4088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4343..4646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4647..4850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4801..4821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(4873..4948,1..5)
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 ccggtcgcca ccatggcgcg taaggtcgat ctcacctcct gcgatcgcga gccgatccac
       61 atccccggca gcattcagcc gtgcggctgc ctgctagcct gcgacgcgca ggcggtgcgg
      121 atcacgcgca ttacggaaaa tgccggcgcg ttctttggac gcgaaactcc gcgggtcggt
      181 gagctactcg ccgattactt cggcgagacc gaagcccatg cgctgcgcaa cgcactggcg
      241 cagtcctccg atccaaagcg accggcgctg atcttcggtt ggcgcgacgg cctgaccggc
      301 cgcaccttcg acatctcgct gcatcgccat gacggtacat cgatcatcga gttcgagcct
      361 gcggcggccg aacaggccga caatccgctg cggctgacgc ggcagatcat cgcgcgcacc
      421 aaagaactga agtcgctcga agagatggcc gcacgggtgc cgcgctatct gcaggcgatg
      481 ctcggctatc accgcgtgat gttgtaccgc ttcgcggacg acggctccgg caaagtgatc
      541 ggcgaggcga agcgcagcga cctcgagagc tttctcggtc agcactttcc ggcgtcgctg
      601 gtcccgcagc aggcgcggct actgtacttg aagaacgcga tccgcgtggt ctcggattcg
      661 cgcggcatca gcagccggat cgtgcccgag cacgacgcct ccggcgccgc gcttgatctg
      721 tcgttcgcgc acctgcgcag catctcgcct atccatctcg aatttctgcg gaacatgggc
      781 gtcagcgcct cgatgtcgct gtcgatcatc attgacggca cgctatgggg attgatcatc
      841 tgtcatcatt acgagccgcg tgccgtgccg atggcgcagc gcgtcgcggc cgaaatgttc
      901 gccgacttct tatcgctgca cttcaccgcc gcccaccacc aacgctaagc ggccgcgact
      961 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     1021 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     1081 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     1141 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     1201 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     1261 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     1321 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     1381 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     1441 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     1501 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     1561 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     1621 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     1681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     1801 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     1861 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     1921 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     1981 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2041 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2101 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     2161 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2221 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2281 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2341 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2401 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2461 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2521 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     2581 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     2641 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     2701 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     2761 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     2821 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     2881 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     2941 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3001 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3061 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3121 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     3181 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     3241 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     3301 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     3361 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     3421 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3481 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3721 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4201 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4261 taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc
     4321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4621 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     4681 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4741 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4801 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4861 accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc gaattctgca
     4921 gtcgacggta ccgcgggccc gggatcca
//
