LOCUS       sequence-321578-        5956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses murine IFN gamma in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5956)
  TITLE     Expression plasmid for production of murine IFN gamma
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5956)
  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: 
            "Unpublished"
COMMENT     Sequence Label: mu-IFNg-pCIneo
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1660..1781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1671..1690)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1725..1744
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1967..2422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2054..2073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2264..2285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2539..2896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2539..2730
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2562..2582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2747..2882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2947..3741
                     /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(3001..3020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3611..3630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3805..3853
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3894..3913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4013..4035
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4073..4091)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4159..4263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4264..4332,4333..5124)
                     /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(4482..4501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5295..5883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5784..5803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagagga tccaccgcgg ccaggaccat gaacgctaca cactgcatct
     1141 tggctttgca gctcttcctc atggctgttt ctggctgtta ctgccacggc acagtcattg
     1201 aaagcctaga aagtctgaat aactatttta actcaagtgg catagatgtg gaagaaaaga
     1261 gtctcttctt ggatatctgg aggaactggc aaaaggatgg tgacatgaaa atcctgcaga
     1321 gccagattat ctctttctac ctcagactct ttgaagtctt gaaagacaat caggccatca
     1381 gcaacaacat aagcgtcatt gaatcacacc tgattactac cttcttcagc aacagcaagg
     1441 cgaaaaagga tgcattcatg agtattgcca agtttgaggt caacaaccca caggtccagc
     1501 gccaagcatt caatgagctc atccgagtgg tccaccagct gttgccggaa tccagcctca
     1561 ggaagcggaa aaggagtcgc tgctgattcg tcgcgactct agagtcgacc cgggcggccg
     1621 cttcccttta gtgagggtta atgcttcgag cagacatgat aagatacatt gatgagtttg
     1681 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     1741 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     1801 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     1861 acaaatgtgg taaaatccga taaggatcga tccgggctgg cgtaatagcg aagaggcccg
     1921 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc
     1981 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2041 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2101 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2161 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2221 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2281 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2341 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2401 aaaatattaa cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     2461 tttcacaccg catacgcgga tctgcgcagc accatggcct gaaataacct ctgaaagagg
     2521 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2581 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2641 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2701 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2761 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2821 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2881 cctaggcttt tgcaaaaagc ttgattcttc tgacacaaca gtctcgaact taaggctaga
     2941 gccaccatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3001 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3061 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3121 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3181 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3241 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3301 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3361 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3421 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     3481 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3541 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3601 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3661 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3721 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3781 ccaacctgcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     3841 ggttttttgt gtgaatcgat agcgataagg atccgcgtat ggtgcactct cagtacaatc
     3901 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     3961 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4021 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4081 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4141 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4201 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4261 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4321 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     4381 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     4441 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4501 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4561 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4621 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4681 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4741 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4801 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4861 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4921 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4981 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5041 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5101 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5161 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5221 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5281 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     5341 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     5401 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     5461 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5521 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5581 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5641 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5701 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5761 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5821 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5881 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5941 atggctcgac agatct
//
