LOCUS       sequence-377743-        5631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mNeonGreen and C-terminally fused PDT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5631)
  AUTHORS   Sakkos JK, Hernandez-Ortiz S, Osteryoung KW, Ducat DC
  TITLE     Orthogonal Degron System for Controlled Protein Degradation in 
            Cyanobacteria.
  JOURNAL   ACS Synth Biol. 2021 Jul 16;10(7):1667-1681. doi: 
            10.1021/acssynbio.1c00035. Epub 2021 Jul 7.
  PUBMED    34232633
REFERENCE   2  (bases 1 to 5631)
  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"; doi: 
            "10.1021/acssynbio.1c00035"; journalName: "ACS Synth Biol"; date: 
            "2021-07-16- 16"; volume: "10"; issue: "7"; pages: "1667-1681"
COMMENT     Sequence Label: pHN1_lacUV5-mNG-PDT
FEATURES             Location/Qualifiers
     source          1..5631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1335..1421
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1513..1540
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(1573..1578,1579..1596,1597..1603)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1611..1627
                     /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        2456..2557
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2558..3217
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2624..2643)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        3282..3359
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3360..4442
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(3379..3398)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(join(5424..5631,1..381))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5504..5523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
       61 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      421 tctagttcag ccagctcgtc gtgatgtcga aacccaagcc accctcagag gtgaaggccg
      481 cttcgagcac atcgggaagc gtgtcgacga cggtgctcgg tgcctcgggt aagagccaga
      541 tagatgcggt gacgttcacc gtcgcgatcg tggcgggaac cacggtcact gcgtccgtta
      601 caactcgcac gtcgtcggcc agcacgactg actcgacggc ttcgattagc ccgggagagg
      661 caaggccatc aggctcggta gacagaatgc tgattaacac ctcgccggga gcagggctgc
      721 tcacagccgc gtccttcacc cgtgggtcag ccgtcagcgc ttgatagcgg taccaggccg
      781 caccgcctgc ggtcgagctg cccttgatcc gctcgatggt gcgatcgcga agctcgccat
      841 ccggctcatt cggctgccgc accacgccgt agaacgcggc aaggttgtcg aggtcggggc
      901 cgtttgagta gcgaagcagt gtcgcaagaa gtgcgtcgtt gatccgctga cgcaggatca
      961 gctcgcgggc tgcgcagacc tccagcagct tgatgaccgg gtcggattcg aggatggcgg
     1021 tgtagctggc gtcacgcgat cgcaggtcgt cgatcaagtc ctgcaggatc agttcaaagt
     1081 ccagcgcttc gatgatggtg ggcgcgggaa tcgtagcaaa gtcaagaacg gtcatgagac
     1141 gactaagccc tccagcgtga tacgcctgcc ctcggggatg tagtagccga tcaggttcag
     1201 ctcgacttga ccagctgcgc tggctgagac gatgcgaacc ttctccagct tcagccgtgg
     1261 ctcccagcga tcgagcgctt cagctgtggc cgccaccagg tcaacgatga gggactggtt
     1321 gatcggctag tcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta
     1381 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     1441 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     1501 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta ctctagtttc
     1561 gaaaccccag gcttgacact ttatgcttcc ggctcgtata atgtgtggaa ttgtgagcgg
     1621 ataacaattt cacacaagga ggaaaaacat atggtcagca aaggtgaaga agacaatatg
     1681 gcgagcctgc cggcgacgca tgaactgcac attttcggta gcatcaacgg tgtggacttc
     1741 gatatggttg gccagggtac cggcaacccg aatgacggtt atgaagaact gaatctgaaa
     1801 agtacgaagg gcgatctgca attttccccg tggattctgg tcccgcatat cggttacggc
     1861 ttccaccagt acctgccgta tccggatggt atgagcccgt ttcaagcggc catggtggac
     1921 ggttctggct atcaggttca tcgtaccatg caattcgaag atggcgcgag tctgaccgtg
     1981 aactatcgct acacgtatga aggttcacac attaaaggcg aagcgcaggt caaaggtacc
     2041 ggttttccgg cggatggtcc ggtgatgacc aactcactga cggcagctga ttggtgccgt
     2101 tcgaaaaaga cctatccgaa tgacaaaacc attatcagca cgtttaagtg gtcttacacc
     2161 acgggtaacg gcaaacgtta tcgcagcacc gcacgtacca cgtacacgtt cgctaaaccg
     2221 atggcggcca actacctgaa aaatcagccg atgtatgttt ttcgcaaaac cgaactgaaa
     2281 cattccaaga cggaactgaa cttcaaggaa tggcaaaagg cgtttacgga tgtgatgggt
     2341 atggacgaac tgtacaaggc tgctaataaa aacgaagaga acacaaacga agttccgacg
     2401 tttatgctga atgctggcca agcaaaccga cgacgggtct aaaagcgtag gatccgatcg
     2461 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     2521 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     2581 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     2641 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     2701 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     2761 tgctcatccg gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     2821 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     2881 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     2941 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3001 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3061 ccccgttttc actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     3121 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     3181 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     3241 cccttgaatt cctactagtc gaagcggcat gcatttacgt tgacaccatc gaatggtgca
     3301 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     3361 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     3421 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     3481 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     3541 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     3601 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     3661 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     3721 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     3781 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     3841 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     3901 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     3961 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     4021 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     4081 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     4141 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     4201 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     4261 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     4321 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     4381 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4441 gatcccacag ccgccagttc cgctggcggc attttaactt tctttaataa tctagtgaca
     4501 agccggggca gacgtgagcc gtagtcccgt cgccagacgc gggtgcccac gggcgtcgtc
     4561 aggatgtccg taattgactg ccggaggtgg tcaatgccct tcagctcctt gccactgtca
     4621 cggctcatgc ctcgggtcat tagtcgcccg ctccggtatc ttcactggct tcgatgattg
     4681 ccgccccgca gctgcagagg tcaccgatcc gagcagtcgg cctctggttg gtaaagaccg
     4741 tgcgactgcc ggtgatgatc gtgttcaagc catgcagggg gcaggcgtga aggtcatcct
     4801 ttcgggccac ggggcggctg ttgacgaagg tgtcgtcgct cccggtgatg atgatcccgc
     4861 cgtgatcggt cacgtcgttt agtcgagcga tgcctggcgt cgtagtcacg ggtttaggtc
     4921 aatacgactt gcggtcactg taacgttgcc ctcggcggtc acgttaacgt cgccttgggc
     4981 ttcgacttgc gcctcctgca caaggatcac aatccgtcct tgggctgcgg tgaggtcgat
     5041 cttgtactca tgcgcttcgc ggtcgtactg gatgattgag tcatcctcga actgcgtctt
     5101 ttggatcgtt tctttgtcct cgatctgggg gtagtcagtc gagaacgcgc cgggcatcgc
     5161 gaagccctga ctgatctcgc cggagggggc catcacgacg acggcctcac cgacctcggg
     5221 cgcccaccag aaccgatcct tgcccgctcg ctgcgtgagc cacggaatcc agtcagtgag
     5281 gagcagcgcc tcgccgctct cctcgtcttc ctcgatcgcg acacggatca gccccttggg
     5341 atagtcagcc tcggctaccc tgcctacgcg gagcaagttg ccgtgacgcc gactgtctcg
     5401 agtctagtcg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg t
//
