LOCUS       sequence-63498-e        4963 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 4963)
  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 4963)
  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: piRFP682-N1
FEATURES             Location/Qualifiers
     source          1..4963
                     /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..963
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 682 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP682
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGVVIAEDRCAEVESKLGLHYPASAVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPCHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    1085..1206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1122..1141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1176..1195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1213..1668)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1350..1371)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1562..1581
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1695..1799
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1801..2158
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1801..1992
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1824..1844)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2009..2144
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2193..2987
                     /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             2193..2987
                     /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(2247..2266)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2857..2876
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3175..3194)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3219..3266
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3595..4183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4084..4103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4358..4661
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4662..4865
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4816..4836
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(4888..4963,1..5)
                     /label=MCS
                     /note="multiple cloning site"
ORIGIN
        1 ccggtcgcca ccatggcgga aggatccgtc gccaggcagc ctgacctctt gacctgcgac
       61 gatgagccga tccatatccc cggtgccatc caaccgcatg gactgctgct cgccctcgcc
      121 gccgacatga cgatcgttgc cggcagcgac aaccttcccg aactcaccgg actggcgatc
      181 ggcgccctga tcggccgctc tgcggccgat gtcttcgact cggagacgca caaccgtctg
      241 acgatcgcct tggccgagcc cggggcggcc gtcggagcac cgatcactgt cggcttcacg
      301 atgcgaaagg acgcaggctt catcggctcc tggcatcgcc atgatcagct catcttcctc
      361 gagctcgagc ctccccagcg ggacgtcgcc gagccgcagg cgttcttccg ccgcaccaac
      421 agcgccatcc gccgcctgca ggccgccgaa accttggaaa gcgcctgcgc cgccgcggcg
      481 caagaggtgc ggaagattac cggcttcgat cgggtgatga tctatcgctt cgcctccgac
      541 ttcagcgggg tggtgatcgc agaggatcga tgcgccgagg tcgagtcaaa actaggcctg
      601 cactatcctg cctcagcggt gccggcgcag gcccgtcggc tctataccat caacccggta
      661 cggatcattc ccgatatcaa ttatcggccg gtgccggtca ccccagacct caatccggtc
      721 accgggcggc cgattgatct tagcttcgcc atcctgcgca gcgtctcgcc ctgccatttg
      781 gagttcatgc gcaacatagg catgcacggc acgatgtcga tctcgatttt gcgcggcgag
      841 cgactgtggg gattgatcgt ttgccatcac cgaacgccgt actacgtcga tctcgatggc
      901 cgccaagcct gcgagctagt cgcccaggtt ctggcctggc agatcggcgt gatggaagag
      961 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1021 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1081 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1201 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     1261 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     1321 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     1381 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     1441 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     1501 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     1561 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     1621 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     1681 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     1741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     1801 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     1861 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     1921 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1981 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2041 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2101 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     2161 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     2221 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2281 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2341 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     2401 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2461 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2521 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2581 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     2641 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     2701 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     2761 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     2821 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     2881 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     2941 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3001 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3061 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3121 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     3181 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     3241 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3301 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3361 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3421 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     3481 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3541 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3601 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3661 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3721 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3781 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3841 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3901 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3961 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4021 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4081 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4141 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4201 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4261 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     4321 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     4381 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     4441 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     4501 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     4561 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     4621 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     4681 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     4741 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     4801 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     4861 gagctggttt agtgaaccgt cagatccgct agcgctaccg gactcagatc tcgagctcaa
     4921 gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat cca
//
