LOCUS       sequence-173860-        6627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HIF-1a sequence without the bHLH domain fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6627)
  TITLE     See lab plasmids
REFERENCE   2  (bases 1 to 6627)
  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: pg-nucODD-EGFP
FEATURES             Location/Qualifiers
     source          1..6627
                     /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(2424..2448)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2507..2516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2513..2515,2516..2518,2519..3232)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2513..2515,2516..2518,2519..3229)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2558..2579)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2819..2838)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3166..3187
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3355..3476
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3392..3411)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3446..3465
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3483..3938)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3620..3641)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3832..3851
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3965..4069
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4071..4428
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4071..4262
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4094..4114)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4279..4414
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4463..5257
                     /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             4463..5257
                     /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(4517..4536)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5127..5146
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5445..5464)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5489..5536
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5865..6453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6354..6373
                     /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 ccatgacctg
      601 cttggtgctg atttgtgaac ccattcctca cccatcaaat attgaaattc ctttagatag
      661 caagactttc ctcagtcgac acagcctgga tatgaaattt tcttattgtg atgaaagaat
      721 taccgaattg atgggatatg agccagaaga acttttaggc cgctcaattt atgaatatta
      781 tcatgctttg gactctgatc atctgaccaa aactcatcat gatatgttta ctaaaggaca
      841 agtcaccaca ggacagtaca ggatgcttgc caaaagaggt ggatatgtct gggttgaaac
      901 tcaagcaact gtcatatata acaccaagaa ttctcaacca cagtgcattg tatgtgtgaa
      961 ttacgttgtg agtggtatta ttcagcacga cttgattttc tcccttcaac aaacagaatg
     1021 tgtccttaaa ccggttgaat cttcagatat gaaaatgact cagctattca ccaaagttga
     1081 atcagaagat acaagtagcc tctttgacaa acttaagaag gaacctgatg ctttaacttt
     1141 gctggcccca gccgctggag acacaatcat atctttagat tttggcagca acgacacaga
     1201 aactgatgac cagcaacttg aggaagtacc attatataat gatgtaatgc tcccctcacc
     1261 caacgaaaaa ttacagaata taaatttggc aatgtctcca ttacccaccg ctgaaacgcc
     1321 aaagccactt cgaagtagtg ctgaccctgc actcaatcaa gaagttgcat taaaattaga
     1381 accaaatcca gagtcactgg aactttcttt taccatgccc cagattcagg atcagacacc
     1441 tagtccttcc gatggaagca ctagacaaag ttcacctgag cctaatagtc ccagtgaata
     1501 ttgtttttat gtggatagtg atatggtcaa tgaattcaag ttggaattgg tagaaaaact
     1561 ttttgctgaa gacacagaag caaagaaccc attttctact caggacacag atttagactt
     1621 ggagatgtta gctccctata tcccaatgga tgatgacttc cagttacgtt ccttcgatca
     1681 gttgtcacca ttagaaagca gttccgcaag ccctgaaagc gcaagtcctc aaagcacagt
     1741 tacagtattc cagcagactc aaatacaaga acctactgct aatgccacca ctaccactgc
     1801 caccactgat gaattaaaaa cagtgacaaa agaccgtatg gaagacatta aaatattgat
     1861 tgcatctcca tctcctaccc acatacataa agaaactact agtgccacat catcaccata
     1921 tagagatact caaagtcgga cagcctcacc aaacagagca ggaaaaggag tcatagaaca
     1981 gacagaaaaa tctcatccaa gaagccctaa cgtgttatct gtcgctttga gtcaaagaac
     2041 tacagttcct gaggaagaac taaatccaaa gatactagct ttgcagaatg ctcagagaaa
     2101 gcgaaaaatg gaacatgatg gttcactttt tcaagcagta ggaattggaa cattattaca
     2161 gcagccagac gatcatgcag ctactacatc actttcttgg aaacgtgtaa aaggatgcaa
     2221 atctagtgaa cagaatggaa tggagcaaaa gacaattatt ttaataccct ctgatttagc
     2281 atgtagactg ctggggcaat caatggatga aagtggatta ccacagctga ccagttatga
     2341 ttgtgaagtt aatgctccta tacaaggcag cagaaaccta ctgcggggtg aagaattact
     2401 cagagctttg gatcaagtta acgacccagc tttcttgtac aaagtggttg atctcaagct
     2461 tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag
     2521 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2581 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     2641 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2701 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2761 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     2821 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2881 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2941 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3001 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3061 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3121 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3181 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg
     3241 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3301 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3361 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3421 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3481 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3541 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3601 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3661 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3721 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3781 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3841 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3901 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     4081 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4201 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4261 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4321 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4381 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4441 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4501 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4561 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4621 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4681 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4741 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4801 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4861 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4921 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4981 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     5041 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     5101 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     5161 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     5221 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     5281 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5341 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5401 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5461 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5521 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5581 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5641 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5701 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6541 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     6601 agttcatagc ccatatatgg agttccg
//
