LOCUS       sequence-350003-        8500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression human RAP containing a C-terminal 6xHis tag in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8500)
  AUTHORS   Bang YJ, Hu Z, Li Y, Gattu S, Ruhn KA, Raj P, Herz J, Hooper LV
  TITLE     Serum amyloid A delivers retinol to intestinal myeloid cells to 
            promote adaptive immunity.
  JOURNAL   Science. 2021 Sep 17;373(6561):eabf9232. doi: 
            10.1126/science.abf9232. Epub 2021 Sep 17.
  PUBMED    34529485
REFERENCE   2  (bases 1 to 8500)
  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: "Science."; 
            date: "2021-09-17"; volume: "373(6561):eabf9232. doi"; pages: " 
            10.1126/science.abf9232. Epub 2021 Sep 17"
COMMENT     Sequence Label: hRAP-pEZT-BM
FEATURES             Location/Qualifiers
     source          1..8500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(14..123)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     enhancer        308..687
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        688..891
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     842..862
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     888..912
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1027..1159
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1142..1161
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1204..1222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2299..2316
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    2362..2950
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2415..2435)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2833..2844)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3092..3226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3129..3148)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3183..3202
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3255..3420
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3604..4058
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3691..3710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3900..3921
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4084..4188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4189..4257,4258..5049)
                     /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(4407..4426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5220..5808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5709..5728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5962..5979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6113..6337
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6404..6937)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7107..7112,7113..7129,7130..7135))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7643..7691
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     7716..7735
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(7776..8486,8487..8489))
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(7818..7839)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     8167..8186
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8426..8447
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 gtcttcgtcg agtgattgta aataaaatgt aatttacagt atagtatttt aattaatata
       61 caaatgattt gataataatt cttatttaac tataatatat tgtgttgggt tgaattaaag
      121 gtccgtatac tccggaatat taataggatc ccggtccgag cgcgcggaat tcaatattgg
      181 ccattagcca tattattcat tggttatata gcataaatca atattggcta ttggccattg
      241 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
      301 ccatgttggc attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      361 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      421 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      481 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      541 gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg
      601 cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc
      661 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt
      721 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      781 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg
      841 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
      901 aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat tgctaacgca
      961 gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact
     1021 gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg
     1081 tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac
     1141 tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt
     1201 acttaatacg actcactata ggctagcatg gcgccgcgga gggtcaggtc gtttctgcgc
     1261 gggctcccgg cgctgctact gctgctgctc ttcctcgggc cctggcccgc tgcgagccac
     1321 ggcggcaagt actcgcggga gaagaaccag cccaagccgt ccccgaaacg cgagtccgga
     1381 gaggagttcc gcatggagaa gttgaaccag ctgtgggaga aggcccagcg actgcatctt
     1441 cctcccgtga ggctggccga gctccacgct gatctgaaga tacaggagag ggacgaactc
     1501 gcctggaaga aactaaagct tgacggcttg gacgaagatg gggagaagga agcgagactc
     1561 atacgcaacc tcaatgtcat cttggccaag tatggtctgg acggaaagaa ggacgctcgg
     1621 caggtgacca gcaactccct cagtggcacc caggaagacg ggctggatga ccccaggctg
     1681 gaaaagctgt ggcacaaggc gaagacctct gggaaattct ccggcgaaga actggacaag
     1741 ctctggcggg agttcctgca tcacaaagag aaagttcacg agtacaacgt cctgctggag
     1801 accctgagca ggaccgaaga aatccacgag aacgtcatta gcccctcgga cctgagcgac
     1861 atcaagggca gcgtcctgca cagcaggcac acggagctga aggagaagct gcgcagcatc
     1921 aaccagggcc tggaccgcct gcgcagggtc agccaccagg gctacagcac tgaggctgag
     1981 ttcgaggagc ccagggtgat tgacctgtgg gacctggcgc agtccgccaa cctcacggac
     2041 aaggagctgg aggcgttccg ggaggagctc aagcacttcg aagccaaaat cgagaagcac
     2101 aaccactacc agaagcagct ggagattgcg cacgagaagc tgaggcacgc agagagcgtg
     2161 ggcgacggcg agcgtgtgag ccgcagccgc gagaagcacg ccctgctgga ggggcggacc
     2221 aaggagctgg gctacacggt gaagaagcat ctgcaggacc tgtccggcag gatctccaga
     2281 gctcggcaca acgaactcca tcatcaccat caccattgac tcgaggcggc cgcgactgca
     2341 gggatccgag gtaccaagct taatcaacct ctggattaca aaatttgtga aagattgact
     2401 ggtattctta actatgttgc tccttttacg ctatgtgaat acgctgcttt aatgcctttg
     2461 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2521 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2581 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2641 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2701 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2761 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2821 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2881 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2941 gcctccccgc ctgcagtctg acaagcttgt cgagaagtac tagaggatca taatcagcca
     3001 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3061 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgcttga
     3241 gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     3301 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     3361 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     3421 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
     3481 aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt
     3541 cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat
     3601 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3661 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3721 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3781 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3841 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
     3901 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3961 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4021 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4081 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4141 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4201 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4261 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4321 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4381 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4441 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4501 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4561 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4621 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4681 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4741 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4801 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4861 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4921 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4981 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5041 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5101 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5161 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5221 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5281 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5341 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5401 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5461 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5521 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5581 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5641 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5701 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5761 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5821 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5881 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     5941 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     6001 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca gccgcgtaac
     6061 ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg
     6121 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     6181 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     6241 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     6301 gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga
     6361 acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg
     6421 ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg
     6481 cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct
     6541 catgcttgag gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga
     6601 ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc agaacgtaag
     6661 ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac
     6721 tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt
     6781 ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc
     6841 cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac
     6901 tgccctgctg cgtaacatcg ttgctgctgc gtaacatcaa acatcgaccc acggcgtaac
     6961 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     7021 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     7081 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg
     7141 ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
     7201 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc
     7261 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc
     7321 ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
     7381 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta
     7441 gctatagttc tagtggttgg ctacgtaccc gtagtggcta tggcagggct tgccgccccg
     7501 acgttggctg cgagccctgg gccttcaccc gaacttgggg gttggggtgg ggaaaaggaa
     7561 gaaacgcggg cgtattggtc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     7621 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt tattctgtct
     7681 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag
     7741 cctcccccat ctcccggttc cgcatgctat gcatccttgt acagctcgtc catgccgaga
     7801 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     7861 tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     7921 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
     7981 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     8041 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     8101 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     8161 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     8221 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     8281 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     8341 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     8401 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     8461 ccggtgaaca gctcctcgcc cttgctcacc gggtgatcaa
//
