LOCUS       sequence-357775-        6685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of 3UTR-HLA-A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6685)
  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"
COMMENT     Sequence Label: pCIneo-RLuc-3UTR-HLA-A_AD
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        305..516
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     467..487
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     513..537
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          677..809
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     792..811
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        854..872
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      876..885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             882..1817
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2176..2297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2187..2206)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2241..2260
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2483..2938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2570..2589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2780..2801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3055..3412
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3055..3246
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3078..3098)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3263..3398
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3463..4257
                     /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(3517..3536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4127..4146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4321..4369
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4410..4429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4529..4551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4589..4607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4675..4779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4780..4848,4849..5640)
                     /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(4998..5017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5811..6399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6300..6319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(6610..6685,1..304)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactgcga tcgcccgccc cgttgacgca aatgggcggt
      481 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcact
      541 agaagcttta ttgcggtagt ttatcacagt taaattgcta acgcagtcag tgcttctgac
      601 acaacagtct cgaacttaag ctgcagtgac tctcttaagg tagccttgca gaagttggtc
      661 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      721 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      781 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
      841 aggctagagt acttaatacg actcactata ggctagccac catggcttcc aaggtgtacg
      901 accccgagca acgcaaacgc atgatcactg ggcctcagtg gtgggctcgc tgcaagcaaa
      961 tgaacgtgct ggactccttc atcaactact atgattccga gaagcacgcc gagaacgccg
     1021 tgatttttct gcatggtaac gctgcctcca gctacctgtg gaggcacgtc gtgcctcaca
     1081 tcgagcccgt ggctagatgc atcatccctg atctgatcgg aatgggtaag tccggcaaga
     1141 gcgggaatgg ctcatatcgc ctcctggatc actacaagta cctcaccgct tggttcgagc
     1201 tgctgaacct tccaaagaaa atcatctttg tgggccacga ctggggggct tgtctggcct
     1261 ttcactactc ctacgagcac caagacaaga tcaaggccat cgtccatgct gagagtgtcg
     1321 tggacgtgat cgagtcctgg gacgagtggc ctgacatcga ggaggatatc gccctgatca
     1381 agagcgaaga gggcgagaaa atggtgcttg agaataactt cttcgtcgag accatgctcc
     1441 caagcaagat catgcggaaa ctggagcctg aggagttcgc tgcctacctg gagccattca
     1501 aggagaaggg cgaggttaga cggcctaccc tctcctggcc tcgcgagatc cctctcgtta
     1561 agggaggcaa gcccgacgtc gtccagattg tccgcaacta caacgcctac cttcgggcca
     1621 gcgacgatct gcctaagatg ttcatcgagt ccgaccctgg gttcttttcc aacgctattg
     1681 tcgagggagc taagaagttc cctaacaccg agttcgtgaa ggtgaagggc ctccacttca
     1741 gccaggagga cgctccagat gaaatgggta agtacatcaa gagcttcgtg gagcgcgtgc
     1801 tgaagaacga gcagtaattc tagatgtgag acagctgcct tgtgtgggac tgagaggcaa
     1861 gagttgttcc tgcccttccc tttgtgactt gaagaaccct gactttgttt ctgcaaaggc
     1921 acctgcatgt gtctgtgttc gtgtaggcat aatgtgagga ggtggggaga ccaccccacc
     1981 cccatgtcca ccatgaccct cttcccacgc tgacctgtgc tccctcccca atcatctttc
     2041 ctgttccaga gaggtggggc tgaggtgtct ccatctctgt ctcaacttca tggtgcactg
     2101 agctgtaact tcttccttcc ctattaaaag cggccgcttc cctttagtga gggttaatgc
     2161 ttcgagcaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     2221 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     2281 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     2341 agatgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atccgataag
     2401 gatcgatccg ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2461 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2881 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     2941 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg
     3001 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
     3061 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     3121 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     3181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     3241 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     3301 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     3361 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga
     3421 ttcttctgac acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga
     3481 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3541 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3601 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3661 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3721 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3781 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3841 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3901 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3961 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     4021 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4081 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4141 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4201 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4261 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc
     4321 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg
     4381 ataaggatcc gcgtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4441 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4501 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4561 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4621 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4681 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4741 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat cttcaatatt
     6481 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     6541 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     6601 cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     6661 ttcatagccc atatatggag ttccg
//
