LOCUS       sequence-173862-        7221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminally Clover-tagged human HIF-1a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7221)
  TITLE     See lab plasmids
REFERENCE   2  (bases 1 to 7221)
  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: HIF-1alpha-Clover
FEATURES             Location/Qualifiers
     source          1..7221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    563..587
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3072..3096)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3188..3209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3796..3817
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3949..4070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3986..4005)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4040..4059
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4077..4532)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4214..4235)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4426..4445
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4559..4663
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4665..5022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4665..4856
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4688..4708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4873..5008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5057..5851
                     /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             5057..5851
                     /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(5111..5130)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5721..5740
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6039..6058)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6083..6130
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6459..7047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6948..6967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagat caacaagttt gtacaaaaaa gcaggcttca ccatggaggg
      601 cgccggcggc gcgaacgaca agaaaaagat aagttctgaa cgtcgaaaag aaaagtctcg
      661 agatgcagcc agatctcggc gaagtaaaga atctgaagtt ttttatgagt ttgctcatca
      721 gttgccactt ccacataatg tgagttcgca tcttgataag gcctctgtga tgaggcttac
      781 catcagctat ttgcgtgtga ggaaacttct ggatgctggt gatttggata ttgaagatga
      841 catgaaagca caggtgaatt gcttttattt gaaagccttg gatggttttg ttatggttct
      901 cacagatgat ggtgacatga tttacatttc tgataatgtg aacaaataca tgggattaac
      961 tcagtttgaa ctaactggac acagtgtgtt tgattttact catccatgtg accatgagga
     1021 aatgagagaa atgcttacac acagaaatgg ccttgtgaaa aagggtaaag aacaaaacac
     1081 acagcgaagc ttttttctca gaatgaagtg taccctaact agccgaggaa gaactatgaa
     1141 cataaagtct gcaacatgga aggtattgca ctgcacaggc cacattcacg tatatgatac
     1201 caacagtaac caacctcagt gtgggtataa gaaaccacct atgacctgct tggtgctgat
     1261 ttgtgaaccc attcctcacc catcaaatat tgaaattcct ttagatagca agactttcct
     1321 cagtcgacac agcctggata tgaaattttc ttattgtgat gaaagaatta ccgaattgat
     1381 gggatatgag ccagaagaac ttttaggccg ctcaatttat gaatattatc atgctttgga
     1441 ctctgatcat ctgaccaaaa ctcatcatga tatgtttact aaaggacaag tcaccacagg
     1501 acagtacagg atgcttgcca aaagaggtgg atatgtctgg gttgaaactc aagcaactgt
     1561 catatataac accaagaatt ctcaaccaca gtgcattgta tgtgtgaatt acgttgtgag
     1621 tggtattatt cagcacgact tgattttctc ccttcaacaa acagaatgtg tccttaaacc
     1681 ggttgaatct tcagatatga aaatgactca gctattcacc aaagttgaat cagaagatac
     1741 aagtagcctc tttgacaaac ttaagaagga acctgatgct ttaactttgc tggccccagc
     1801 cgctggagac acaatcatat ctttagattt tggcagcaac gacacagaaa ctgatgacca
     1861 gcaacttgag gaagtaccat tatataatga tgtaatgctc ccctcaccca acgaaaaatt
     1921 acagaatata aatttggcaa tgtctccatt acccaccgct gaaacgccaa agccacttcg
     1981 aagtagtgct gaccctgcac tcaatcaaga agttgcatta aaattagaac caaatccaga
     2041 gtcactggaa ctttctttta ccatgcccca gattcaggat cagacaccta gtccttccga
     2101 tggaagcact agacaaagtt cacctgagcc taatagtccc agtgaatatt gtttttatgt
     2161 ggatagtgat atggtcaatg aattcaagtt ggaattggta gaaaaacttt ttgctgaaga
     2221 cacagaagca aagaacccat tttctactca ggacacagat ttagacttgg agatgttagc
     2281 tccctatatc ccaatggatg atgacttcca gttacgttcc ttcgatcagt tgtcaccatt
     2341 agaaagcagt tccgcaagcc ctgaaagcgc aagtcctcaa agcacagtta cagtattcca
     2401 gcagactcaa atacaagaac ctactgctaa tgccaccact accactgcca ccactgatga
     2461 attaaaaaca gtgacaaaag accgtatgga agacattaaa atattgattg catctccatc
     2521 tcctacccac atacataaag aaactactag tgccacatca tcaccatata gagatactca
     2581 aagtcggaca gcctcaccaa acagagcagg aaaaggagtc atagaacaga cagaaaaatc
     2641 tcatccaaga agccctaacg tgttatctgt cgctttgagt caaagaacta cagttcctga
     2701 ggaagaacta aatccaaaga tactagcttt gcagaatgct cagagaaagc gaaaaatgga
     2761 acatgatggt tcactttttc aagcagtagg aattggaaca ttattacagc agccagacga
     2821 tcatgcagct actacatcac tttcttggaa acgtgtaaaa ggatgcaaat ctagtgaaca
     2881 gaatggaatg gagcaaaaga caattatttt aataccctct gatttagcat gtagactgct
     2941 ggggcaatca atggatgaaa gtggattacc acagctgacc agttatgatt gtgaagttaa
     3001 tgctcctata caaggcagca gaaacctact gcggggtgaa gaattactca gagctttgga
     3061 tcaagttaac gacccagctt tcttgtacaa agtggttgat ctcaagcttc gaattctgca
     3121 gtcgacggta ccgcgggccc ggatggtgag caagggcgag gagctgttca ccggggtggt
     3181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
     3241 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3301 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgg cctgcttcag
     3361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3421 cgtccaggag cgcaccatct ctttcaagga cgacggtacc tacaagaccc gcgccgaggt
     3481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3541 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat
     3601 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagatccgcc acaacgttga
     3661 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3721 cgtgctgctg cccgacaacc actacctgag ccatcagtcc gccctgagca aagaccccaa
     3781 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgcctgag gccgcgactc
     3841 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3901 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     4081 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4141 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4201 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4261 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4321 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4381 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4441 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4501 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4561 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4621 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4681 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4861 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4921 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4981 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     5041 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5101 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5161 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5221 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5281 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5341 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5401 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5461 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5521 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5581 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5641 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5701 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5761 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5821 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5881 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5941 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6001 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     6061 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     6121 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6181 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6241 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6301 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7081 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7141 aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt cattagttca
     7201 tagcccatat atggagttcc g
//
