LOCUS       sequence-181612-        8220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Type II Hexokinase expressed in AS-30D rat hepatoma cell line.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8220)
  AUTHORS   Mathupala SP, Heese C, Pedersen PL
  TITLE     Glucose catabolism in cancer cells. The type II hexokinase promoter 
            contains functionally active response elements for the tumor 
            suppressor p53.
  JOURNAL   J Biol Chem. 1997 Sep 5;272(36):22776-80.
  PUBMED    9278438
REFERENCE   2  (bases 1 to 8220)
  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 Biol 
            Chem."; date: "1997-09-5"; volume: "272(36)"; pages: "22776-80"
COMMENT     Sequence Label: pCI-Neo-HKIImut
FEATURES             Location/Qualifiers
     source          1..8220
                     /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"
     regulatory      1102..1111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3924..4045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3935..3954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3989..4008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      4231..4686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4318..4337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4528..4549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4803..5160
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4803..4994
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4826..4846)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5011..5146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5211..6005
                     /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(5265..5284)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5875..5894
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6069..6117
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(6158..6177)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6277..6299
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6337..6355)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6423..6527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6528..6596,6597..7388)
                     /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"
     rep_origin      7559..8147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8048..8067
                     /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 ctcgagaatt cgccaccatg gtcgcctcgc atatgatcgc ctgcttattc
     1141 acggagctca accaaaacca agtgcagaag gttgaccaat ttctctacca catgcgtctc
     1201 tcagatgaga cccttctgga gatttctagg cggttccgga aggagatgga gaaagggcta
     1261 ggagctacca cgcaccctac agcagctgtg aaaatgttgc ctacctttgt gaggtcaact
     1321 ccggatggga cagaacatgg ggagttcctg gctctggatc ttggaggaac caacttccgt
     1381 gtgctccgag taagggtgac ggacaatggc ctccagagag tggagatgga gaaccagatc
     1441 tacgccatcc ttgaggacat catgcggggc agtggaaccc agctgtttga ccacatcgcc
     1501 gaatgcctgg ccaacttcat ggacaagcta caaatcaaag agaagaagct ccctctgggt
     1561 ttcaccttct cgttcccctg ccaccagaca aaactggatg agagtttttt ggtctcgtgg
     1621 actaaggggt tcaagtccag tggcgtggaa ggcagagatg tggtggacct gatccggaag
     1681 gttatccagc gcagagggga ctttgacatt gacattgtgg ccgtggtgaa tgacacagtt
     1741 gggaccatga tgacttgtgg ctatgatgat cagaactgcg agattggtct cattgtgggc
     1801 actggcagca acgcctgcta catggaggaa atgcgtcata ttgacatggt ggagggagat
     1861 gaggggcgca tgtgcatcaa catggagtgg ggagcctttg gggacgacgg tacactcaat
     1921 gacatccgaa ccgagtttga ccgagagatc gacatgggct cgctgaaccc tgggaagcag
     1981 ctgtttgaga agatgattag cgggatgtac atgggggagc tggtcaggct catcctggtg
     2041 aagatggcca aggcagagct gttgttccaa gggaaactca gcccagaact ccttaccact
     2101 ggctccttcg agaccaaaga tgtctcggat attgaagagg ataaggatgg aatcgagaag
     2161 gcctaccaaa tcctgatgcg cctgggtctg aatccattgc aggaggattg tgtggccacg
     2221 caccgaatct gccagattgt gtccacgcgc tcggccagtc tgtgcgcagc caccctggcc
     2281 gcggtgctgt ggcgaatcaa agagaacaag ggcgaggagc gacttcgctc caccatcggt
     2341 gtcgatggct ccgtctacaa gaaacatccc cattttgcca agcgtctcca taaggcagtg
     2401 aggaggctgg tgcccgactg tgatgtccgc ttcctccgct ctgaggatgg cagcggcaag
     2461 ggggctgcta tggtgacggc ggtggcttac cgtctggctg accaacaccg ggcccgccag
     2521 aagaccctgg agtctctgaa gctgagccac gagcagcttc tggaggttaa gagaagaatg
     2581 aaggtggaaa tggagcaggg tctgagcaag gagacgcatg cggtcgcccc tgtgaagatg
     2641 ctgcccactt acgtgtgtgc cactccagat ggcacagaga aaggagactt cttggccttg
     2701 gatcttggag gaacaaactt ccgggtcctg ctggtgcgtg tgcgtaatgg caagcggagg
     2761 ggcgtggaga tgcataacaa gatctactcc atcccacagg aggttatgca tggcactggg
     2821 gaagagctct tcgaccacat tgtccagtgc attgcggact tcctggagta catgggcatg
     2881 aagggcgtgt ccctgccttt gggtttcaca ttctccttcc cttgccagca gaacagccta
     2941 gaccagagca tcctcctcaa gtggacaaag ggattcaagg catctggctg cgagggtgag
     3001 gatgtggtca ccttgctgaa ggaagcgatt caccggcgag aggagtttga cctggatgtg
     3061 gttgccgtgg tgaatgacac agttgggact atgatgactt gtggctacga agaccctcac
     3121 tgtgaagttg gcctcattgt tggcaccgga agcaacgcct gctacatgga agagatgcgt
     3181 aatgtggagc tggtggacgg agaggaggga cggatgtgtg tcaacatgga gtggggagca
     3241 tttggggaca atggctgcct ggatgacttg cggaccgtgt ttgatgttgc tgtggatgag
     3301 ctttctctca accctggcaa acagaggttc gagaagatga tcagcggcat gtacttggga
     3361 gagattgtgc gcaacattct catcgatttc acgaagcggg ggctgctctt ccgaggccgc
     3421 atctcagagc gcctcaagac aaggggaatc tctgaaacta agttcctgtc tcagatagag
     3481 agcgactgcc tagccctgct acaggttcgt gccatcctgc gccacctagg gctggagagc
     3541 acgtgcgatg acagcatcat cgtgaaggag gtgtgcactg tggttgcccg gcgcgctgca
     3601 cagctctgtg gcgcaggcat ggccgccgta gtggacaaga taagagagaa ccgtgggctg
     3661 gacaacccca aagtgacagt gggcgtggac gggactctgt ataagcttca tcctcacttt
     3721 gccaaggtca tgcatgagac ggtgagagat ctggctccga aatgtgacgt gtccttcctg
     3781 gaatccgagg acggcagtgg gaagggagca gctctcatca ctgccgtggc ctgccgcatc
     3841 cgggaggctg ggcagagata gtctagagtc gacccgggcg gccgcttccc tttagtgagg
     3901 gttaatgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     4021 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     4081 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     4141 ccgataagga tcgatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     4201 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     4261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4441 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4561 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4621 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     4681 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4741 cggatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc
     4801 ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4861 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4921 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4981 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5041 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5101 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5161 aagcttgatt cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac
     5221 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     5281 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5341 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     5401 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5461 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5521 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5581 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5641 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5701 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     5761 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5821 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5881 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5941 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     6001 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     6061 atggccgcaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
     6121 cgatagcgat aaggatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat
     6181 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6241 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6301 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6361 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6541 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6601 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6661 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6721 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6781 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6841 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6901 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     6961 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7021 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7081 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7141 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7201 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7261 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7321 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7381 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7441 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7501 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7561 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7621 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7681 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7741 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7801 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7861 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7921 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     7981 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8041 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8101 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8161 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct
//
