LOCUS       sequence-439386-        6862 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Igbp1-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6862)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 6862)
  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: Igbp1-GFP
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(120..139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      933..1521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1422..1441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1675..1692
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1809..1830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1845..1850,1851..1868,1869..1875)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1883..1899
                     /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     1888..1910
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1907..1923
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1907..1923
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1972..2093
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1983..2002)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2037..2056
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2223..3245)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2223..3245)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(3253..3300)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3432..3451
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3584..3808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3797..3814
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3858..4571,4572..4574,4575..4577))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     4252..4271
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4511..4532
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4590..4610)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5735..5759)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5769..5787
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5790..5808
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5810..6013)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5839..5859)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6014..6393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6565..6584
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6659..6763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6764..6832,6833..6862,1..762)
                     /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 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
       61 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
      121 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
      181 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
      241 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
      301 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
      361 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
      421 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
      481 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
      541 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
      601 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
      661 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
      721 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
      781 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
      841 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
      901 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
      961 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1021 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     1081 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     1141 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     1201 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     1261 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     1321 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     1381 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     1441 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     1501 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     1561 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     1621 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     1681 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     1741 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     1801 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     1861 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     1921 gaccatgatt acgccaagct ctagctagag gtcgacggta tacagacatg ataagataca
     1981 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2041 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttggggtgg
     2101 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     2161 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtagtacg
     2221 tgctattcct ttgccctcgg acgagtgctg gggcgtcggt ttccactatc ggcgagtact
     2281 tctacacagc catcggtcca gacggccgcg cttctgcggg cgatttgtgt acgcccgaca
     2341 gtcccggctc cggatcggac gattgcgtcg catcgaccct gcgcccaagc tgcatcatcg
     2401 aaattgccgt caaccaagct ctgatagagt tggtcaagac caatgcggag catatacgcc
     2461 cggagccgcg gcgatcctgc aagctccgga tgcctccgct cgaagtagcg cgtctgctgc
     2521 tccatacaag ccaaccacgg cctccagaag aagatgttgg cgacctcgta ttgggaatcc
     2581 ccgaacatcg cctcgctcca gtcaatgacc gctgttatgc ggccattgtc cgtcaggaca
     2641 ttgttggagc cgaaatccgc gtgcacgagg tgccggactt cggggcagtc ctcggcccaa
     2701 agcatcagct catcgagagc ctgcgcgacg gacgcactga cggtgtcgtc catcacagtt
     2761 tgccagtgat acacatgggg atcagcaatc gcgcatatga aatcacgcca tgtagtgtat
     2821 tgaccgattc cttgcggtcc gaatgggccg aacccgctcg tctggctaag atcggccgca
     2881 gcgatcgcat ccatggcctc cgcgaccggc tgcagaacag cgggcagttc ggtttcaggc
     2941 aggtcttgca acgtgacacc ctgtgcacgg cgggagatgc aataggtcag gctctcgctg
     3001 aattccccaa tgtcaagcac ttccggaatc gggagcgcgg ccgatgcaaa gtgccgataa
     3061 acataacgat ctttgtagaa accatcggcg cagctattta cccgcaggac atatccacgc
     3121 cctcctacat cgaagctgaa agcacgagat tcttcgccct ccgagagctg catcaggtcg
     3181 gagacgctgt cgaacttttc gatcagaaac ttctcgacag acgtcgcggt gagttcaggc
     3241 tttttggcca aggaagttcc tatactttct agagaatagg aacttcggaa taggaacttc
     3301 taggtacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3361 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3421 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3481 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtgg
     3541 ggataccccc tagagcccca gctggttctt tccgcctcag aagccataga gcccaccgca
     3601 tccccagcat gcctgctatt gtcttcccaa tcctccccct tgctgtcctg ccccacccca
     3661 ccccccagaa tagaatgaca cctactcaga caatgcgatg caatttcctc attttattag
     3721 gaaaggacag tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca
     3781 acagatggct ggcaactaga aggcacagtc gaggctgatc agcgggttta aacgggccct
     3841 ctagactcga gtcactatta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     3901 gtcacgaact ccttaaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3961 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     4021 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     4081 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     4141 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     4201 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     4261 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     4321 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag
     4381 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     4441 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     4501 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     4561 tcgcccttgc tcaccatggt ggtaccttca ccttgaaaat ataaattttc gcggccgcgg
     4621 cccatgttct gtcggttccc atagccccta ggatgggtgt ccttccagtc atcccactcc
     4681 cgggctctgt ggagtgtttg ttcatcatcc tcttcctcct tttcttcttg ttcttcctgt
     4741 tgctgagctg cttttctgaa ttcctctggt gctgccttgg ctattccctg atccggtaat
     4801 gctccatatt tccgatgttg ctcataccag tcactcaccg tcatagttgg cagacttgga
     4861 taaccagctc caaatacttt ggcttgagcc atgttccgag tgagaatgaa gggtttcact
     4921 ggaggcctct cctggcgaga tgagttagaa gttgatgcct ctcttgaaga gtctctttct
     4981 ctcaggatct ttatttcctg gtcaatgctc tcaatctctt ctaagctgat atcaatccac
     5041 ctctgaaggt gaagaagata atattcacga acacgctcat catctgcttg accactttcc
     5101 acagcagatt tcattgcaga caacctatgc tccaactcct tcttctgctt gtatctctgt
     5161 attttagcct gtctttgaga tgccatagca acgagactag gataagccat ggaggaattg
     5221 gcagtgtgat tttcagcaga gttgttcatg gttttgggca gctcaaactc tgccacatga
     5281 tagcaatggc actgagttaa gtagtttata aagtgttctc gagcccgctg caaatgatct
     5341 agacgcttgc tggggttgac ttgtttcatg gtgagggctc cttgaaacgc tggcaccaaa
     5401 aggtacttca ggtcggtgga agcaatctct tccaaatctt catttcggct gaacaagtcg
     5461 agctgcgata acatttcggc agccttctca aggaggtcca agcccttgaa caccttctcc
     5521 tggactatcc gggaaccggc gggttcagtc gccacttcta cttcgtccag taactgtcta
     5581 ccagtttcga acagctcggg gagccgcggc agctgtaact cgtcctcagc agccatgcag
     5641 gcctaagctt aagtttaaac gctagagtcc ggaggctgga tcggtcccgg tgtcttctat
     5701 ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact aaacgagctc
     5761 gtcgacgatc tctatcactg atagggagat ctctatcact gatagggaga gctctgctta
     5821 tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg gagttgttac
     5881 gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg acgtcaatgg
     5941 ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc
     6001 aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa
     6061 agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg tcattgacgt
     6121 caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg gcagtttacc
     6181 gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact atgggaacat
     6241 acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc gggccattta
     6301 ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac cccgtaattg
     6361 attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc cgtacatcgc
     6421 gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg gtcaagcctt
     6481 gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac acacaagcag
     6541 ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     6601 ggggatcggg agatctcccg atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg
     6661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     6721 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     6781 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6841 aacgctggtg aaagtaaaag at
//
