LOCUS       sequence-409037-        6076 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing the mutant S39E of human IF1 fused in C-terminal 
            with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6076)
  TITLE     px458-sgIF1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6076)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pIF1-S39E-GFP
FEATURES             Location/Qualifiers
     source          1..6076
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      20..29
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(26..28,29..31,32..745)
                     /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(26..28,29..31,32..742)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(71..92)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(332..351)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     679..700
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(842..859)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    848..1072
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1118..1546
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1205..1224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1415..1436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1555..1575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1560..1889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1740..1875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1802..1821
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1937..1984
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2003..2377
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2507..2628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2544..2563)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2598..2617
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2677..2693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2677..2693)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2690..2712)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2701..2717
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2725..2731,2732..2749,2750..2755))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2770..2791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2908..2925)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3079..3667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3159..3178)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3838..4629,4630..4698))
                     /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     4461..4480
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4699..4803)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4878..4897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5069..5448
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5449..5652
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5603..5623
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5697..5716
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5697..5715
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggcccgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
       61 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
      121 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
      181 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
      241 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
      301 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
      361 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
      421 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
      481 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat
      541 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
      601 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc
      661 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
      721 cggcatggac gagctgtaca agtaaagcgg ccgccactgt gctggatatc tgcagaattc
      781 caccacactg gactagtgga tccgagctcg gtaccaagct taagtttaaa ccgctgatca
      841 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
      901 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
      961 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     1021 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     1081 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     1141 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1201 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1261 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1321 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1381 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1441 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1501 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     1561 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1621 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     1681 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     1741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     1801 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     1861 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     1921 gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata
     1981 cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc
     2041 gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt
     2101 cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt
     2161 ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga
     2221 gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc
     2281 catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg
     2341 caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc
     2401 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     2461 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     2521 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2581 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     2641 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4801 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     4861 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     4921 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     4981 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     5041 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     5101 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     5161 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     5221 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     5281 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     5341 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     5401 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     5461 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     5521 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     5581 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5641 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     5701 acgactcact atagggagac ccaagctggc tagcgctacc ggactcagat ctcgagatgg
     5761 cagtgacggc gttggcggcg cggacgtggc ttggcgtgtg gggcgtgagg accatgcaag
     5821 cccgaggctt cggctcggat cagtccgaga atgtcgaccg gggcgcgggc gagatccggg
     5881 aagccggtgg ggccttcgga aagagagagc aggctgaaga ggaacgatat ttccgagcac
     5941 agagtagaga acaactggca gctttgaaaa aacaccatga agaagaaatc gttcatcata
     6001 agaaggagat tgagcgtctg cagaaagaaa ttgagcgcca taagcagaag atcaaaatgc
     6061 taaaacatga tgatag
//
