LOCUS       sequence-156018-        6079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  YFP expressed from pJ23119 in the glpK locus with gentamicin 
            resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6079)
  AUTHORS   Gordon GC, Korosh TC, Cameron JC, Markley AL, Begemann MB, Pfleger 
            BF
  TITLE     CRISPR interference as a titratable, trans-acting regulatory tool 
            for metabolic engineering in the cyanobacterium Synechococcus sp. 
            strain PCC 7002.
  JOURNAL   Metab Eng. 2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007.
  PUBMED    27481676
REFERENCE   2  (bases 1 to 6079)
  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: "Metab Eng. 
            2016 Jul 29. pii: S1096-7176(16)30062-3. doi: 
            10.1016/j.ymben.2016.07.007."
COMMENT     Sequence Label: pGCG26
FEATURES             Location/Qualifiers
     source          1..6079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     494..513
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(732..836)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     904..922
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(960..982)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1082..1101
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1295..1317
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1309..1326
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1310..1326
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1944..1978
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     CDS             2030..2059
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             2072..2083
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(2105..2107,2108..2110,2111..2824)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVRFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2150..2171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2411..2430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2758..2779
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2894..2912)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2908..2955
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        join(3129..3134,3135..3151,3152..3157)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3346..3879
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     primer_bind     complement(4789..4805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4789..4805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4802..4824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4813..4829
                     /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(4837..4843,4844..4861,4862..4867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4882..4903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5020..5037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5191..5779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5271..5290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5950..6079,1..662,663..731))
                     /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 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
       61 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      121 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      181 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      241 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      301 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      361 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      421 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      481 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      541 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      601 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      661 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      721 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      781 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      841 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
      901 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
      961 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1021 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1081 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1141 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     1201 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1261 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1321 gccagtgaat tcgagctcgg tacccgggtg aagcgattgg ctatgatcta ccaaagctgg
     1381 cgatttttct gcaaaccgaa ttagaaccac aatttgattt tttaaagccg attaccattg
     1441 ccaacctgaa ggaaatagcg atcgccttta tggaggatgg ccacgaggct tgtcgggtct
     1501 cccattattg tggatcggta ccggatgagc gggcgagctt taatttacgc ttgcggcaat
     1561 atcgccaggc ccagccttgg ctgcgaaatc atctcgatcc tgcaaggggc gatcgcctgc
     1621 accattggtc ggatcaccaa cgcaccattt tctacggcag gcgcaccaat cccgacaccc
     1681 agcagcggct tgtgttagtg gcgcacatgg ccggggctcc gaagaccgtt gagattggca
     1741 aatggctcgc cctggatttg gatcgttggc agttggcgat cgccacaccg actttgaaga
     1801 tcaacaccat ctatgactta gcccaaattc acttgcacaa tggcgaaggt tttctgttat
     1861 ctgaaattcc tccctaaatg atgtcttttg agcctaaaaa cacacttttt tgacctaatt
     1921 taacccattt aaaaaagctc gagttgacag ctagctcagt cctaggtata atgctagcta
     1981 ctagtgcatc gctatcgcgt atactattta cgagacacat accatgggcc atcatcatca
     2041 tcatcatcat catcatcaca gcagcggcca tatcgaaggt cgtcatatgc aggcagcagc
     2101 ctcgatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2161 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2221 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2281 caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     2341 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2401 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaggttcg agggcgacac
     2461 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2521 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2581 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2641 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2701 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2761 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2821 gtaacgaggg cggtgctttg gcaggatccg gctgctaaca aagcccgaaa ggaagctgag
     2881 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2941 ttgacgggtt ttttgtctag acgcatgcat aaaaactgtt gtaattcatt aagcattctg
     3001 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     3061 ttgtcgcctt gcgtataata tttgcccatg gacgcacacc gtggaaacgg atgaaggcac
     3121 gaacccagtt gacataagcc tgttcggttc gtaaactgta atgcaagtag cgtatgcgct
     3181 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     3241 tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc aagcagcaag
     3301 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagc
     3361 aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc aagtatgggc
     3421 atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc tgctcttgat
     3481 cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc ggactccgat
     3541 tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa
     3601 gcggttgttg gcgctctcgc ggcttacgtt ctgcccaggt ttgagcagcc gcgtagtgag
     3661 atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat tgccaccgcg
     3721 ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa
     3781 gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat acgggaagaa
     3841 gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca agccgagatc
     3901 ggcttcccgg ccgcggagtt gttcggtaaa ttgtcacaac gccgccaggt ggcacttttc
     3961 ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc gctggacttc
     4021 ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg ccttggcctg
     4081 catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct cccgaaggtg
     4141 gatccccccc tcttgcctac agcatctccc ccaggggaga attcttcctg tttcaactcc
     4201 ctctaacgta aacccattga atttaaaaaa gactttatga ctgctttact gctccatgac
     4261 caacattatt cccttgatca tgaagccttt ctctcaaccc tcagcaacac agaaaattta
     4321 ctcattattc aagatctaga tggcgtttgc atggggttag tcaaagaccc cttaacccgc
     4381 aaaattgatc ctgactatat ccgcgccaca cgcaagttta gagaccactt ctttgtcctc
     4441 accaacggtg aacatgaagg cagaagggga gtaaatcgca tcgttgaacg ggcatttcgc
     4501 aatgttgaag ccaaagagga aacaagctat ttacctggtt tagcagcagg gggtgtgcaa
     4561 tggcagacag ataatggcca aatttcccat cccggtgtta gccaagcaga actcgatttc
     4621 cttgccacag tgccagattt aattggtcaa agtttaggac aattttttac taaatatgtt
     4681 gatatttttc ccgctgagct tcaacctgag ctgatccatg cttctgtttt agataatctc
     4741 gtttcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     4801 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4861 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4921 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4981 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5041 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5101 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5161 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5221 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5281 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5341 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5401 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5461 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5521 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5581 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5641 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5701 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5761 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5821 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5881 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5941 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6001 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6061 atctggcccc agtgctgca
//
