LOCUS       sequence-372862-        7424 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses IDO2 and GFP (seperately) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7424)
  AUTHORS   Maassen S, Warner H, Ioannidis M, Jansma J, Markus H, El Aidy S, 
            Chiara MD, Chiara JL, Maierhofer L, Weavers H, van den Bogaart G
  TITLE     Mitochondrial interaction of fibrosis-protective 5-methoxy 
            tryptophan enhances collagen uptake by macrophages.
  JOURNAL   Free Radic Biol Med. 2022 Aug 1;188:287-297. doi: 
            10.1016/j.freeradbiomed.2022.06.235. Epub 2022 Jun 23.
  PUBMED    35753585
REFERENCE   2  (bases 1 to 7424)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Free Radic 
            Biol Med."; date: "2022-08-1"; volume: "188:287-297. doi"; pages: " 
            10.1016/j.freeradbiomed.2022.06.235. Epub 2022 Jun 23"
COMMENT     Sequence Label: pcDNA3.1-IDO2-p2a-eGFP
FEATURES             Location/Qualifiers
     source          1..7424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..466
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        467..670
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     621..641
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     715..734
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        715..733
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2068..2124
                     /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             join(2125..2127,2128..2841)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2125..2838
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2167..2188)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2428..2447)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2775..2796
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2867..2884)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2873..3097
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3143..3571
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3230..3249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3440..3461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3580..3600)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3585..3914
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3765..3900
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3827..3846
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3981..4775
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4035..4054)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4645..4664
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4949..5070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4986..5005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5040..5059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5119..5135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5119..5135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5132..5154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5143..5159
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5167..5173,5174..5191,5192..5197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5212..5233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5350..5367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5521..6109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5601..5620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6280..7071,7072..7140))
                     /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     6903..6922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7141..7245)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7320..7339)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
      121 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      181 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      241 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      301 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      361 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      421 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      481 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      541 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      601 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      661 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      721 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgg taccgagctc
      781 ggatccatgc tgcacttcca ctactacgac accagcaaca agatcatgga gccccaccgc
      841 cccaacgtga agaccgccgt gcccctgagc ctggagagct accacatcag cgaggagtac
      901 ggcttcctgc tgcccgacag cctgaaggag ctgcccgacc actaccgccc ctggatggag
      961 atcgccaaca agctgcccca gctgatcgac gcccaccagc tgcaggccca cgtggacaag
     1021 atgcccctgc tgagctgcca gttcctgaag ggccaccgcg agcagcgcct ggcccacctg
     1081 gtgctgagct tcctgaccat gggctacgtg tggcaggagg gcgaggccca gcccgccgag
     1141 gtgctgcccc gcaacctggc cctgcccttc gtggaggtga gccgcaacct gggcctgccc
     1201 cccatcctgg tgcacagcga cctggtgctg accaactgga ccaagaagga ccccgacggc
     1261 ttcctggaga tcggcaacct ggagaccatc atcagcttcc ccggcggcga gagcctgcac
     1321 ggcttcatcc tggtgaccgc cctggtggag aaggaggccg tgcccggcat caaggccctg
     1381 gtgcaggcca ccaacgccat cctgcagccc aaccaggagg ccctgctgca ggccctgcag
     1441 cgcctgcgcc tgagcatcca ggacatcacc aagaccctgg gccagatgca cgactacgtg
     1501 gaccccgaca tcttctacgc cggcatccgc atcttcctga gcggctggaa ggacaacccc
     1561 gccatgcccg ccggcctgat gtacgagggc gtgagccagg agcccctgaa gtacagcggc
     1621 ggcagcgccg cccagagcac cgtgctgcac gccttcgacg agttcctggg catccgccac
     1681 agcaaggaga gcggcgactt cctgtaccgc atgcgcgact acatgccccc cagccacaag
     1741 gccttcatcg aggacatcca cagcgccccc agcctgcgcg actacatcct gagcagcggc
     1801 caggaccacc tgctgaccgc ctacaaccag tgcgtgcagg ccctggccga gctgcgcagc
     1861 taccacatca ccatggtgac caagtacctg atcaccgccg ccgccaaggc caagcacggc
     1921 aagcccaacc acctgcccgg ccccccccag gccctgaagg accgcggcac cggcggcacc
     1981 gccgtgatga gcttcctgaa gagcgtgcgc gacaagaccc tggagagcat cctgcacccc
     2041 cgcggcctcg agtctagagg aagcggagct actaacttca gcctgctgaa gcaggctgga
     2101 gacgtggagg agaaccctgg acctatgagc aagggcgagg agctgttcac cggggtggtg
     2161 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     2221 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2281 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2341 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2401 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2461 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2521 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2581 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2641 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2701 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2761 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactcacggc
     2821 atggacgagc tgtacaagta agggcccgtt taaacccgct gatcagcctc gactgtgcct
     2881 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     2941 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3001 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3061 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3121 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3181 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3241 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3301 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3361 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3421 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3481 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3541 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3781 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3841 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3901 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     3961 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4021 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4081 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4141 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4201 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4261 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4321 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4381 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4441 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4501 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4561 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4621 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4681 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4741 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4801 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4861 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4921 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     4981 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5041 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5101 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5161 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5221 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5281 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5341 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5401 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5461 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5521 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5581 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5641 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5701 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5761 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5821 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5881 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5941 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6001 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6061 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6121 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6181 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6241 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6301 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6361 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6421 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6481 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6541 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6601 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6661 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6721 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6781 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6841 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6901 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6961 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7021 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7081 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7141 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7201 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7261 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     7321 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     7381 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggca
//
