LOCUS       sequence-422681-        5642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of mouse IL2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5642)
  AUTHORS   Heller L, Pottinger C, Jaroszeski MJ, Gilbert R, Heller R
  TITLE     In vivo electroporation of plasmids encoding GM-CSF or interleukin-2
            into existing B16 melanomas combined with electrochemotherapy 
            induces long-term antitumour immunity.
  JOURNAL   Melanoma Res. 2000 Dec;10(6):577-83.
  PUBMED    11198480
REFERENCE   2  (bases 1 to 5642)
  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: "Melanoma 
            Res."; date: "2000-12"; volume: "10(6)"; pages: "577-83"
COMMENT     Sequence Label: pCR3.1-mIL2
FEATURES             Location/Qualifiers
     source          1..5642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..588
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     539..559
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1384..1401)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1390..1614
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1746..2334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1826..1845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2663..2710
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2735..2754
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2942..3736)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(2942..3736)
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3053..3072)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3663..3682
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3771..4128)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3785..3920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3937..4128
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4085..4105
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4188..4979,4980..5048))
                     /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     4811..4830
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5049..5153)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5180..5635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5267..5286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5477..5498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      421 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      481 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      541 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      601 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      661 gctggctagc gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt
      721 ggaattcggc ttccttgcta atcactcctc acagtgacct caagtcctgc aggcatgtac
      781 agcatgcagc tcgcatcctg tgtcacattg acacttgtgc tccttgtcaa cagcgcaccc
      841 acttcaagct ccacttcaag ctctacagcg gaagcacagc agcagcagca gcagcagcag
      901 cagcagcagc agcacctgga gcagctgttg atggacctac aggagctcct gagcaggatg
      961 gagaattaca ggaacctgaa actccccagg atgctcacct tcaaatttta cttgcccaag
     1021 caggccacag aattgaaaga tcttcagtgc ctagaagatg aacttggacc tctgcggcat
     1081 gttctggatt tgactcaaag caaaagcttt caattggaag atgctgagaa tttcatcagc
     1141 aatatcagag taactgttgt aaaactaaag ggctctgaca acacatttga gtgccaattc
     1201 gatgatgagt cagcaactgt ggtggacttt ctgaggagat ggatagcctt ctgtcaaagc
     1261 atcatctcaa caagccctca ataactatgt acctcctgct tacaacacaa gccgaattct
     1321 gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat
     1381 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1441 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     1501 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     1561 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     1621 aggcggaaag aaccagtggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1681 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1741 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1801 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1861 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1921 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1981 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2041 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2101 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2161 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2221 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2281 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2341 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2401 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2461 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     2521 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     2581 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     2641 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     2701 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     2761 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     2821 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     2881 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     2941 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     3001 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     3061 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     3121 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     3181 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     3241 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     3301 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     3361 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     3421 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     3481 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     3541 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     3601 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     3661 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     3721 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag
     3781 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     3841 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     3901 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     3961 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     4021 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     4081 ccacacccta actgacacac attccacagc tggttctttc cgcctcagga ctcttccttt
     4141 ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4201 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4261 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4321 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4381 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4441 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4501 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4561 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4621 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4681 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4741 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4801 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4861 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4921 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4981 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5041 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5101 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5161 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5221 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5521 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5581 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttacgcg
     5641 cg
//
