LOCUS       sequence-170219-        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"
     regulatory      73..82
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(79..81,82..84,85..798)
                     /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(79..81,82..84,85..795)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(124..145)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(385..404)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     732..753
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    921..1042
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(958..977)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1012..1031
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1049..1504)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1186..1207)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1398..1417
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1531..1635
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1637..1994
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1637..1828
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1660..1680)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1845..1980
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2029..2823
                     /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             2029..2823
                     /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(2083..2102)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2693..2712
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3011..3030)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3055..3102
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3431..4019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3920..3939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4194..4497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4498..4701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4652..4672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4756..4780
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(join(6617..6627,1..14))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 ttgtacaaag tggttgatct caagcttcga attctgcagt cgacggtacc gcgggcccgg
       61 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      121 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      181 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      241 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      301 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      361 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      421 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      481 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      541 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      601 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      661 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
      721 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
      781 gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg
      841 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
      901 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
      961 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1021 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     1081 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     1141 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     1201 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     1261 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     1321 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     1381 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     1441 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     1501 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     1561 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1621 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     1681 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     1741 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     1801 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     1861 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     1921 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     1981 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     2041 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2101 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2161 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     2221 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     2281 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     2341 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     2401 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     2461 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     2521 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     2581 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     2641 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     2701 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     2761 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     2821 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2881 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2941 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     3001 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3061 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3121 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3181 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3241 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     3301 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3361 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3421 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3481 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3541 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     3601 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3661 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3721 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3781 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3841 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3901 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3961 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4021 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4081 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat
     4141 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     4201 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     4261 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     4321 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     4381 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     4441 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     4501 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     4561 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     4621 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     4681 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
     4741 cagatctcga gatcaacaag tttgtacaaa aaagcaggct tcaccatgac ctgcttggtg
     4801 ctgatttgtg aacccattcc tcacccatca aatattgaaa ttcctttaga tagcaagact
     4861 ttcctcagtc gacacagcct ggatatgaaa ttttcttatt gtgatgaaag aattaccgaa
     4921 ttgatgggat atgagccaga agaactttta ggccgctcaa tttatgaata ttatcatgct
     4981 ttggactctg atcatctgac caaaactcat catgatatgt ttactaaagg acaagtcacc
     5041 acaggacagt acaggatgct tgccaaaaga ggtggatatg tctgggttga aactcaagca
     5101 actgtcatat ataacaccaa gaattctcaa ccacagtgca ttgtatgtgt gaattacgtt
     5161 gtgagtggta ttattcagca cgacttgatt ttctcccttc aacaaacaga atgtgtcctt
     5221 aaaccggttg aatcttcaga tatgaaaatg actcagctat tcaccaaagt tgaatcagaa
     5281 gatacaagta gcctctttga caaacttaag aaggaacctg atgctttaac tttgctggcc
     5341 ccagccgctg gagacacaat catatcttta gattttggca gcaacgacac agaaactgat
     5401 gaccagcaac ttgaggaagt accattatat aatgatgtaa tgctcccctc acccaacgaa
     5461 aaattacaga atataaattt ggcaatgtct ccattaccca ccgctgaaac gccaaagcca
     5521 cttcgaagta gtgctgaccc tgcactcaat caagaagttg cattaaaatt agaaccaaat
     5581 ccagagtcac tggaactttc ttttaccatg ccccagattc aggatcagac acctagtcct
     5641 tccgatggaa gcactagaca aagttcacct gagcctaata gtcccagtga atattgtttt
     5701 tatgtggata gtgatatggt caatgaattc aagttggaat tggtagaaaa actttttgct
     5761 gaagacacag aagcaaagaa cccattttct actcaggaca cagatttaga cttggagatg
     5821 ttagctccct atatcccaat ggatgatgac ttccagttac gttccttcga tcagttgtca
     5881 ccattagaaa gcagttccgc aagccctgaa agcgcaagtc ctcaaagcac agttacagta
     5941 ttccagcaga ctcaaataca agaacctact gctaatgcca ccactaccac tgccaccact
     6001 gatgaattaa aaacagtgac aaaagaccgt atggaagaca ttaaaatatt gattgcatct
     6061 ccatctccta cccacataca taaagaaact actagtgcca catcatcacc atatagagat
     6121 actcaaagtc ggacagcctc accaaacaga gcaggaaaag gagtcataga acagacagaa
     6181 aaatctcatc caagaagccc taacgtgtta tctgtcgctt tgagtcaaag aactacagtt
     6241 cctgaggaag aactaaatcc aaagatacta gctttgcaga atgctcagag aaagcgaaaa
     6301 atggaacatg atggttcact ttttcaagca gtaggaattg gaacattatt acagcagcca
     6361 gacgatcatg cagctactac atcactttct tggaaacgtg taaaaggatg caaatctagt
     6421 gaacagaatg gaatggagca aaagacaatt attttaatac cctctgattt agcatgtaga
     6481 ctgctggggc aatcaatgga tgaaagtgga ttaccacagc tgaccagtta tgattgtgaa
     6541 gttaatgctc ctatacaagg cagcagaaac ctactgcagg gtgaagaatt actcagagct
     6601 ttggatcaag ttaacgaccc agctttc
//
