LOCUS       sequence-204871-        9076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSIN-cytosFD-P2A-cytosFNR, expressing Synechococcus sp Fd and FNR, 
            under EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9076)
  AUTHORS   Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Hu VJ, Chao SS, 
            Hsu A, Pham V, Naghavian L, Dozier LE, Patrick GN, Coleman TP
  TITLE     Biosynthesis of Orthogonal Molecules Using Ferredoxin and 
            Ferredoxin-NADP(+) Reductase Systems Enables Genetically Encoded 
            PhyB Optogenetics.
  JOURNAL   ACS Synth Biol. 2018 Feb 16;7(2):706-717. doi: 
            10.1021/acssynbio.7b00413. Epub 2018 Jan 24.
  PUBMED    29301067
REFERENCE   2  (bases 1 to 9076)
  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"; doi: 
            "10.1021/acssynbio.7b00413"; journalName: "ACS Synth Biol"; date: 
            "2018-02-16- 16"; volume: "7"; issue: "2"; pages: "706-717"
COMMENT     Sequence Label: pPKm-241
FEATURES             Location/Qualifiers
     source          1..9076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          232..1170
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1127..1147
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1273..1282
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1282..1323
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1636..1692
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             1708..1725
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2955..2975)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2960..3289
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3140..3275
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3299..3898
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3299..3318)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3795..3815
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4191..4424
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4575..4592)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4581..4805
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4848..4857
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4901..5005
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5006..5074,5075..5866)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5224..5243)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6037..6625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6526..6545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             6784..7417
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7464..7589
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8082..8315
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8500..8544
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8831..8948
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
       61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
      121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
      181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
      241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
      301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
      781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
      841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
      901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
      961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattgatcc ttcgaactag
     1201 aattcgaagc ttgagctcga gatctgagtc cggtagcgct agttcgaagg atccgctagc
     1261 acattcgaag ccgccaccat gggcaagccc atccccaacc ccctgctggg cctggactcc
     1321 accgttaaca tggctactta caaagtgacc ctgattacac ccgatgggga ggtgtcctac
     1381 gatgcccccg acgatgagta tattctggac agcgccggcg atgcagggta cgatctgcca
     1441 gcttcttgca gagctggcgc ctgcagtacc tgcgcaggga aaatcgtgag cggaacagtg
     1501 gatcagagtg aacagtcctt cctggacgat gaccagatcg aggccggcta tgtgctgaca
     1561 tgtattgcct acccacagtc cgatgtgacc attgagacca acaaggagga agagctgtac
     1621 gtgtacggat ctggggccac aaatttttcc ctgctgaaac aggctgggga cgtggaggaa
     1681 aatccaggcc ccgcatccgg atctatgcat catcaccacc accatgttaa catgtatgga
     1741 atcacctcaa ccgccaactc aaccggaaat cagtcctacg ctaatcggct gttcatctac
     1801 gaagtggtcg ggctgggagg cgacggacgg aacgagaata gcctggtgcg caagtccggg
     1861 accacattca tcactgtccc ttacgcacga atgaaccagg aaatgcagag gattaccaag
     1921 ctgggcggaa aaatcgtgtc tattcgacca gccgaggatg ccgctcagat cgtcagtgaa
     1981 ggacagagct ccgctcaggc atcagcccag agccctatgg ccagcagcac aaagattgtg
     2041 caccccaaaa ctaccgacac tagcgtgcct gtcaacatct accgccctaa gaccccattc
     2101 ctgggcaaat gcattgagaa ttatgaactg gtggatgagg ggggctcagg aactgtgagg
     2161 catgtcacct ttgacatcag cgagggggat ctgagatacc tggaagggca gtccattggc
     2221 atcattcccc ctggagagga caaaaacggg aagcctcaca aactgcgcct gtattccatc
     2281 gcctctaccc gacatggcga catggaggat aataagaccg tgagcctgtg cgtgaggcag
     2341 ctggaatacc aggaccccga gtctggcgaa accgtgtacg gcgtgtgcag cacatacctg
     2401 tgcaacctgc cagtgggaac agacgatgtc aagatcactg gaccagtggg gaaagagatg
     2461 ctgctgcccg acgatgaaga cgcaacagtg gtcatgctgg ccacagggac tggcattgcc
     2521 ccattcaggg cttttctgtg gagaatgttt aaggagcagc acgaagatta taagttcaaa
     2581 gggaaggcct ggctgatctt tggcgtgccc tacactgcta atattctgta taaggacgat
     2641 ttcgagaaaa tggcagccga aaaccctgac aacttcaggc tgacttacgc tatctcccgg
     2701 gagcagaaga ccgcagacgg agggaaagtg tacgtccaga gtcgggtgtc agaatatgcc
     2761 gatgagctgt ttgaaatgat ccagaagccc aacacacacg tgtacatgtg cggcctgaaa
     2821 ggaatgcagc cacccattga cgagaccttt acagctgagg cagaaaagcg cggcctgaat
     2881 tgggaggaaa tgaggaggtc tatgaaaaaa gagcatcggt ggcatgtgga ggtgtattga
     2941 taagtcgacc ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3001 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3061 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3121 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3181 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3241 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaccat
     3301 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg
     3361 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg
     3421 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     3481 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     3541 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     3601 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     3661 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     3721 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     3781 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     3841 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg
     3901 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgataaa
     3961 ataaaagatt ttatttagtc tccagaaaaa gggggcggcc gctcgagacc tagaaaaaca
     4021 tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc
     4081 acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac
     4141 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     4201 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     4261 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4321 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     4381 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     4441 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg
     4501 acattataat agatttagca ggaattgaac taggagtgga gcacacaggc aaagttctag
     4561 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4621 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4681 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4741 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4801 tatggcttct gaggcggaaa gaaccagctg ggggcgcgcc cctcgaggcc gccatggtca
     4861 tagctgtttg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4921 atttttctaa atacattcaa atatgtatcc gctcacgaga caataaccct gataaatgct
     4981 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5041 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5101 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5161 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5221 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5281 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5341 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5401 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5461 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5521 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5581 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5641 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5701 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5761 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5821 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5881 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5941 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6001 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6061 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttctggcc
     6661 ttttgcacgg accgccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc
     6721 ctctgaatta ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt
     6781 agttggaagg gctaattcac tcccaaagaa gacaagatat ccttgatctg tggatctacc
     6841 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6901 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6961 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     7021 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     7081 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     7141 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     7201 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     7261 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7321 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7381 ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac agggacttga
     7441 aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct gaagcgcgca
     7501 cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact agcggaggct
     7561 agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt agatcgcgat
     7621 gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa catatagtat
     7681 gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa acatcagaag
     7741 gctgtagaca aatactggga cagctacaac catcccttca gacaggatca gaagaactta
     7801 gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata gagataaaag
     7861 acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag accaccgcac
     7921 agcaaccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     7981 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     8041 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     8101 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     8161 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     8221 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     8281 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     8341 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     8401 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     8461 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     8521 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     8581 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     8641 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     8701 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     8761 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccacaa atggcagtat
     8821 tcatccacaa ttttaaaaga aaggggggga ttggggggta cagtgcaggg gaaagaatag
     8881 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     8941 aaaattttcg ggtttattac agggacagca gagatccact ttggatcgat aagctttgca
     9001 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
     9061 gaaaggagtg ggaatt
//
