LOCUS       sequence-468364-        8023 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Tool for genetically encoded fluorescent biosensor of cellular iron 
            microenvironment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  TITLE     FEOX
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8023)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: FEOX-PBCS-TBF2- MC2
FEATURES             Location/Qualifiers
     source          1..8023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(106..125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      919..1507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1408..1427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1661..1678
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1795..1816
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1831..1836,1837..1854,1855..1861)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1869..1885
                     /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)."
     primer_bind     1874..1896
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1893..1909
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1893..1909
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1930..1948
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2097..2131
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    2498..2722
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2850..3560)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3128..3147)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3397..3415
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3557..3566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3610..4120)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3673..3691)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     polyA_signal    4127..4182
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(4219..4929)
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      5424..5433
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5465..6643)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5474..6412
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(5497..5517)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     repeat_region   7013..7075
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7141..7160)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7142..7160)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7167..7184)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7167..7183)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7176..7198)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7325..7780
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7412..7431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7622..7643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7806..7910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7911..7979,7980..8023,1..748)
                     /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"
ORIGIN
        1 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
       61 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
      301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
      361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
      421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
      481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
      541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
      601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
      661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
      721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
      781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
      841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1561 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1621 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     1681 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     1741 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1801 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1861 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1921 caagctcgaa attaaccctc actaaaggga acaaaagctg gtacctcgcg cgacttggtt
     1981 tgccattctt tagcgcgcgt cgcgtcacac agcttggcca caatgtggtt tttgtcaaac
     2041 gaagattcta tgacgtgttt aaagtttagg tcgagtaaag cgcaaatctt ttttaaccct
     2101 agaaagatag tctgcgtaaa attgacgcat gcattcttga aatattgctc tctctttcta
     2161 aatagcgcga atccgtcgct gtgcatttag gacatctcag tcgccgcttg gagctcccgt
     2221 gaggcgtgct tgtcaatgcg gtaagtgtca ctgattttga actataacga ccgcgtgagt
     2281 caaaatgacg catgattatc ttttacgtga cttttaagat ttaactcata cgataattat
     2341 attgttattt catgttctac ttacgtgata acttattata tatatatttt cttgttatag
     2401 atatcgtgac taatatataa taaaatgggt agttctttag acgatgagca tatcctctct
     2461 gctcttctgc aaagcgatga cgagcttgtt ggctagacca tagagcccac cgcatcccca
     2521 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     2581 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     2641 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     2701 ggctggcaac tagaaggcac agagacagat cttgtacaat cgatcgccgg agaccatatt
     2761 atccctgcgg ggagggggcc agcgggggaa ggagggggag gccaggccgg gaagagaggg
     2821 aggaagacgg gcggtttaga tctaagcttc tacttgtaca gctcgtccat gccgccggtg
     2881 gagtggcggc cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag
     2941 gtgatgtcca acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc
     3001 ttgtaggtgg tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc
     3061 ttgatctcgc ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag
     3121 cccatggtct tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc
     3181 accttgtaga tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg
     3241 ccgtcctcga agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag
     3301 tagtcgggga tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg
     3361 gacaggatgt cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc
     3421 tgggtgccct cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc
     3481 acggagccct ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc
     3541 tcctcgccct tgctcaccat ggtggcgaat cgatgtcgac aaacagcgct accggtgtcg
     3601 acggatctgc tggggagaga ggtcggtgat tcggtcaacg agggagccga ctgccgacgt
     3661 gcgctccgga ggcttgcaga atgcggaaca ccgcgcgggc aggaacaggg cccacactac
     3721 cgccccacac cccgcctccc gcaccgcccc ttcccggccg ctgctctcgg cgcgccctgc
     3781 tgagcagccg ctattggcca cagcccatcg cggtcggcgc gctgccattg ctccctggcg
     3841 ctgtccgtct gcgagggtac tagtgagacg tgcggcttcc gtttgtcacg tccggcacgc
     3901 cgcgaaccgc aaggaacctt cccgacttag gggcggagca ggaagcgtcg ccggggggcc
     3961 cacaagggta gcggcgaaga tccgggtgac gctgcgaacg gacgtgaaga atgtgcgaga
     4021 cccagggtcg gcgccgctgc gtttcccgga accacgccca gagcagccgc gtccctgcgc
     4081 aaacccaggg ctgccttgga aaaggcgcaa ccccaacccc ctcgagtgag agacacaaaa
     4141 aattccaaca cactattgca atgaaaataa atttccttta ttcaattggt ttcccggggt
     4201 taacgtttaa actacctaat taagcttgtg ccccagtttg ctagggaggt cgcagtatct
     4261 ggccactgcc acctcgtgct gctcgacgta ggtctcgttg ttggcctcct tgattctttc
     4321 cagtctgtag tccacatagt agacgccagg catcttgagg ttcttagcgg gtttcttgga
     4381 tctatatgtg gtcttggcgt ttgcgatcag atggctcccg cccacgagct tcagggccat
     4441 gtcgtttctg ccttccaggc cgccgtcagc ggggtacagc gtctcggtga aggcctccca
     4501 gccgagtgtt ttcttctgca tcacagggcc gttggatgtg aagttcaccc ctctgatctt
     4561 gacgttgtag atgaggcagc cgtcctggag gctggtgtcc tgggtagcgg tcagcacgcc
     4621 cccgtcttcg tatgtggtga ctctctccca tgtgaagccc tcagggaagg actgcttgaa
     4681 gaagtcgggg atgccctggg tgtggttgat gaaggtcttg ctgccgtaga ggaagctagt
     4741 agccaggatg tcgaaggcga aggggagagg gccgccctcg accaccttga ttctcatggt
     4801 ctgggtgccc tcgtagggct tgccttcgcc ctcggatgtg cacttgaagt gatggttgtc
     4861 cacggtgccc tccatgtaca gcttcatgtg catgttctcc ttaatcagct cttcgccctt
     4921 agacaccata agtaggatcc gtttctgaga gcagtgttgt gcaatcactt tctttttaat
     4981 atccttaagc tcttcatagg taaaatattc cattaacatg ggctgaaaaa cctcctcttc
     5041 ctctttcatg tgaggaagaa aatctcttgt aaaagcctcc aatctctctt tcagttgttt
     5101 tgcataattt aactgttcat attcattctt aacattcttc agtccctttt caaagaggct
     5161 aagcatctcg gagagtttat tgtcagaatg tacattataa atggtctggc tgcgttgttg
     5221 aagcaaacca ataatgtatt cattttcaat ctgctcatgc attttgaact ccttgaaagt
     5281 agcatacaaa gactgcagaa gagcacggaa atcgttgttg ttggaaaaat tggttttaga
     5341 aagcttgtcg cagtagagcc ccaccagctg cttcatccgc cagtgtgggg cggtgaagac
     5401 gtccacttct tcaggaaagg gcgccatggc ggcaaacaaa cgcggccgcg acgtccccct
     5461 atagtcacga cacctgaaat ggaagaaaaa aactttgaac cactgtctga ggcttgagaa
     5521 tgaaccaaga tccaaactca aaaagggcaa attccaagga gaattacatc aagtgccaag
     5581 ctggcctaac ttcagtctcc acccactcag tgtggggaaa ctccatcgca taaaacccct
     5641 ccccccaacc taaagacgac gtactccaaa agctcgagaa ctaatcgagg tgcctggacg
     5701 gcgcccggta ctccgtggag tcacatgaag cgacggctga ggacggaaag gcccttttcc
     5761 tttgtgtggg tgactcaccc gcccgctctc ccgagcgccg cgtcctccat tttgagctcc
     5821 ctgcagcagg gccgggaagc ggccatcttt ccgctcacgc aactggtgcc gaccgggcca
     5881 gccttgccgc ccagggcggg gcgatacacg gcggcgcgag gccaggcacc agagcaggcc
     5941 ggccagcttg agactacccc cgtccgattc tcggtggccg cgctcgcagg ccccgcctcg
     6001 ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc cgcggcccca aaaaccgaaa
     6061 taccagtgtg cagatcttgg cccgcattta caagactatc ttgccagaaa aaaagcgtcg
     6121 cagcaggtca tcaaaaattt taaatggcta gagacttatc gaaagcagcg agacaggcgc
     6181 gaaggtgcca ccagattcgc acgcggcggc cccagcgccc aggccaggcc tcaactcaag
     6241 cacgaggcga aggggctcct taagcgcaag gcctcgaact ctcccaccca cttccaaccc
     6301 gaagctcggg atcaagaatc acgtactgca gccaggtgga agtaattcaa ggcacgcaag
     6361 ggccataacc cgtaaagagg ccaggcccgc gggaaccaca cacggcactt acctgtgttc
     6421 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
     6481 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
     6541 cgcgccacct tctctaggca ccggttcaat tgccgacccc tccccccaac ttctcgggga
     6601 ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gcctcacgtg tacaggggct
     6661 agcgaattct taattaacga gagcataata ttgatatgtg ccaaagttgt ttctgactga
     6721 ctaataagta taatttgttt ctattatgta taggttaagc taattactta ttttataata
     6781 caacatgact gtttttaaag tacaaaataa gtttattttt gtaaaagaga gaatgtttaa
     6841 aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat aaataaataa
     6901 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
     6961 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
     7021 aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaaa
     7081 taatagtttc taattttttt attattcagc ctgctgtcgt gaataccgag ctccaattcg
     7141 ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa
     7201 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     7261 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     7321 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     7381 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     7441 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     7501 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     7561 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     7621 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     7681 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     7741 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     7801 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     7861 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     7921 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     7981 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gtt
//
