LOCUS       sequence-453695-        7502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP-tagged human Chromogranin A in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7502)
  AUTHORS   Ioannidis M, van Dijk H, Muntjewerff EM, Chirasani VR, Warner H, van
            den Dool W, Grijpstra P, Bianchi F, Mahata SK, van den Bogaart G
  TITLE     Inflammation Promotes Proteolytic Processing of the Prohormone 
            Chromogranin A by Macrophages.
  JOURNAL   J Endocr Soc. 2025 May 15;9(7):bvaf090. doi: 10.1210/jendso/bvaf090.
            eCollection 2025 Jul.
  PUBMED    40458085
REFERENCE   2  (bases 1 to 7502)
  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: "J Endocr 
            Soc."; date: "2025-05-15"; volume: "9(7):bvaf090. doi"; pages: " 
            10.1210/jendso/bvaf090. eCollection 2025 Jul"
COMMENT     Sequence Label: CHGA-eGFP
FEATURES             Location/Qualifiers
     source          1..7502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2345..2347,2348..3061)
                     /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             2345..3058
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2387..2408)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2648..2667)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2995..3016
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3093..3110)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3099..3323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3369..3797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3456..3475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3666..3687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3806..3826)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3811..4140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3991..4126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4053..4072
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4207..5001
                     /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(4261..4280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4871..4890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5175..5296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5212..5231)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5266..5285
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5345..5361)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5345..5361)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5358..5380)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5369..5385
                     /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(5393..5399,5400..5417,5418..5423))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5438..5459
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5576..5593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5747..6335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5827..5846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6506..7297,7298..7366))
                     /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     7129..7148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7367..7471)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttatgc gctccgccgc tgtcctggct cttctgctct gcgccgggca
      961 agtcactgcg ctccctgtga acagccctat gaataaaggg gataccgagg tgatgaaatg
     1021 catcgttgag gtcatctccg acacactttc caagcccagc cccatgcctg tcagccagga
     1081 atgttttgag acactccgag gagatgaacg gatcctttcc attctgagac atcagaattt
     1141 actgaaggag ctccaagacc tcgctctcca aggcgccaag gagagggcac atcagcagaa
     1201 gaaacacagc ggttttgaag atgaactctc agaggttctt gagaaccaga gcagccaggc
     1261 cgagctgaaa gaggcggtgg aagagccatc atccaaggat gttatggaga aaagagagga
     1321 ttccaaggag gcagagaaaa gtggtgaagc cacagacgga gccaggcccc aggccctccc
     1381 ggagcccatg caggagtcca aggctgaggg gaacaatcag gcccctgggg aggaagagga
     1441 ggaggaggag gaggccacca acacccaccc tccagccagc ctccccagcc agaaataccc
     1501 aggcccacag gccgaggggg acagtgaggg cctctctcag ggtctggtgg acagagagaa
     1561 gggcctgagt gcagagccag ggtggcaggc aaagagagaa gaggaggagg aggaggagga
     1621 ggaggctgag gctggagagg aggctgtccc cgaggaagaa ggccccactg tagtgctgaa
     1681 cccccacccg agccttggct acaaggagat ccggaaaggc gagagtcggt cggaggctct
     1741 ggctgtggat ggagctggga agcctggggc tgaggaggct caggaccccg aagggaaggg
     1801 agaacaggag cactcccagc agaaagagga ggaggaggag atggcagtgg tcccgcaagg
     1861 cctcttccgg ggtgggaaga gcggagagct ggagcaggag gaggagcggc tctccaagga
     1921 gtgggaggac tccaaacgct ggagcaagat ggaccagctg gccaaggagc tgacggctga
     1981 gaagcggctg gaggggcagg aggaggagga ggacaaccgg gacagttcca tgaagctctc
     2041 cttccgggcc cgggcctacg gcttcagggg ccctgggccg cagctgcgac gaggctggag
     2101 gccatcctcc cgggaggaca gccttgaggc gggcctgccc ctccaggtcc gaggctaccc
     2161 cgaggagaag aaagaggagg agggcagcgc aaaccgcaga ccagaggacc aggagctgga
     2221 gagcctgtcg gccattgaag cagagctgga gaaagtggcc caccagctgc aggcactacg
     2281 gcggggcgga tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg
     2341 ccgcatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2401 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2461 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2521 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2581 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2641 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2701 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2761 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2821 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2881 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2941 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     3001 cctgctggag ttcgtgaccg ccgccgggat cactcacggc atggacgagc tgtacaagta
     3061 atctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3121 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3181 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3241 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3301 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3361 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3421 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3481 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3541 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3601 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3661 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3721 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3781 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3901 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3961 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4021 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4081 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4141 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4201 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4261 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4321 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4381 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4441 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4501 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4561 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4621 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4681 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4741 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4801 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4861 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4921 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4981 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5041 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5101 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5161 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5221 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5281 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5341 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5401 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5461 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5521 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5581 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7501 tc
//
