LOCUS       sequence-288135-        6354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suicide plasmid for integration of PlacUV5-cscB and lacI into 
            Synechococcus sp. UTEX 2973 Neutral site III.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6354)
  AUTHORS   Lin PC, Zhang F, Pakrasi HB
  TITLE     Enhanced production of sucrose in the fast-growing cyanobacterium 
            Synechococcus elongatus UTEX 2973.
  JOURNAL   Sci Rep. 2020 Jan 15;10(1):390. doi: 10.1038/s41598-019-57319-5.
  PUBMED    31942010
REFERENCE   2  (bases 1 to 6354)
  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: "Sci Rep."; 
            date: "2020-01-15"; volume: "10(1):390. doi"; pages: " 
            10.1038/s41598-019-57319-5"
COMMENT     Sequence Label: pSL3005
FEATURES             Location/Qualifiers
     source          1..6354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      425..511
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      603..630
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        join(663..668,669..686,687..693)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    701..717
                     /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     complement(2222..2241)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2233..3048
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIYNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2306..2325)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        3095..3172
                     /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             3173..4255
                     /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(3192..3211)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(5237..5825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5317..5336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgaagcagt gtcgcaagaa gtgcgtcgtt gatccgctga cgcaggatca gctcgcgggc
       61 tgcgcagacc tccagcagct tgatgaccgg gtcggattcg aggatggcgg tgtagctggc
      121 gtcacgcgat cgcaggtcgt cgatcaagtc ctgcaggatc agttcaaagt ccagcgcttc
      181 gatgatggtg ggcgcgggaa tcgtagcaaa gtcaagaacg gtcatgagac gactaagccc
      241 tccagcgtga tacgcctgcc ctcggggatg tagtagccga tcaggttcag ctcgacttga
      301 ccagctgcgc tggctgagac gatgcgaacc ttctccagct tcagccgtgg ctcccagcga
      361 tcgagcgctt cagctgtggc cgccaccagg tcaacgatga gggactggtt gatcggctag
      421 tcatcaaata aaacgaaagg ctcagtcgga agactgggcc tttcgtttta tctgttgttt
      481 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
      541 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
      601 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta ctctagtttc gaaaccccag
      661 gcttgacact ttatgcttcc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
      721 cacacaagga ggaaaaacat atgtctagaa tggcactgaa tattccattc agaaatgcgt
      781 actatcgttt tgcatccagt tactcatttc tcttttttat ttcctggtcg ctgtggtggt
      841 cgttatacgc tatttggctg aaaggacatc tagggttgac agggacggaa ttaggtacac
      901 tttattcggt caaccagttt accagcattc tatttatgat gttctacggc atcgttcagg
      961 ataaactcgg tctgaagaaa ccgctcatct ggtgtatgag tttcatcctg gtcttgaccg
     1021 gaccgtttat gatttacgtt tatgaaccgt tactgcaaag caatttttct gtaggtctaa
     1081 ttctgggggc gctctttttt ggcctggggt atctggcggg atgcggtttg cttgacagct
     1141 tcaccgaaaa aatggcgcga aattttcatt tcgaatatgg aacagcgcgc gcctggggat
     1201 cttttggcta tgctattggc gcgttctttg ccggcatatt ttttagtatc agtccccata
     1261 tcaacttctg gttggtctcg ctatttggcg ctgtatttat gatgatcaac atgtgtttta
     1321 aagataagga tcaccagtgc gtagcggcgg atgcgggagg ggtaaaaaaa gaggatttta
     1381 tcgcagtttt caaggatcga aacttctggg ttttcgtcat atttattgtt gggacgtggt
     1441 ctttctataa catttttgat caacaacttt ttcctgtctt ttatgcaggt ttattcgaat
     1501 cacacgatgt aggaacgcgc ctgtatggtt atctcaactc attccaggtg gtactcgaag
     1561 cgctgtgcat ggcgattatt ccgttctttg tgaatcgggt agggccaaaa aatgcattac
     1621 ttatcggtgt tgtgattatg gcgttgcgta tcctttcctg cgcgctgttc gttaacccct
     1681 ggattatttc attagtgaag ctgttacatg ccattgaggt tccactttgt gtcatatccg
     1741 tcttcaaata cagcgtggca aactttgata agcgcctgtc gtcgacgatc tttctgattg
     1801 gttttcaaat tgccagttcg cttgggattg tgctgctttc aacgccgact gggatactct
     1861 ttgaccacgc aggctaccag acagttttct tcgcaatttc gggtattgtc tgcctgatgt
     1921 tgctatttgg cattttcttc ctgagtaaaa aacgcgagca aatagttatg gaaacgcctg
     1981 taccttcagc aatatagact agtagcggcc gcgtaggatc cgatcggcac gtaagaggtt
     2041 ccaactttca ccataatgaa ataagatcac taccgggcgt cgagctgatc cttcaactca
     2101 gcaaaagttc gatttattca acaaagccac gttgtgtctc aaaatctctg atgttacatt
     2161 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     2221 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctccaggcc gcgattaaat
     2281 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
     2341 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
     2401 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
     2461 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
     2521 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
     2581 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
     2641 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
     2701 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
     2761 caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt cgtcactcat
     2821 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
     2881 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     2941 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat ttataatcct
     3001 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     3061 aattggttgt aacactggca gagcggtacc atgggacacc atcgaatggt gcaaaacctt
     3121 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     3181 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     3241 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     3301 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     3361 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     3421 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     3481 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     3541 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     3601 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     3661 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     3721 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     3781 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     3841 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     3901 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     3961 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     4021 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     4081 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     4141 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     4201 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgatccca
     4261 cagccgccag ttccgctggc ggcattttaa ctttctttaa taatctagtg acaagccggg
     4321 gcagacgtga gccgtagtcc cgtcgccaga cgcgggtgcc cacgggcgtc gtcaggatgt
     4381 ccgtaattga ctgccggagg tggtcaatgc ccttcagctc cttgccactg tcacggctca
     4441 tgcctcgggt cattagtcgc ccgctccggt atcttcactg gcttcgatga ttgccgcccc
     4501 gcagctgcag aggtcaccga tccgagcagt cggcctctgg ttggtaaaga ccgtgcgact
     4561 gccggtgatg atcgtgttca agccatgcag ggggcaggcg tgaaggtcat cctttcgggc
     4621 cacggggcgg ctgttgacga aggtgtcgtc gctcccggtg atgatgatcc cgccgtgatc
     4681 ggtcacgtcg tttagtcgag cgatgcctgg cgtcgtagtc acgggtttag gtcaatacga
     4741 cttgcggtca ctgtaacgtt gccctcggcg gtcacgttaa cgtcgccttg ggcttcgact
     4801 tgcgcctcct gcacaaggat cacaatccgt ccttgggctg cggtgaggtc gatcttgtac
     4861 tcatgcgctt cgcggtcgta ctggatgatt gagtcatcct cgaactgcgt cttttggatc
     4921 gtttctttgt cctcgatctg ggggtagtca gtcgagaacg cgccgggcat cgcgaagccc
     4981 tgactgatct cgccggaggg ggccatcacg acgacggcct caccgacctc gggcgcccac
     5041 cagaaccgat ccttgcccgc tcgctgcgtg agccacggaa tccagtcagt gaggagcagc
     5101 gcctcgccgc tctcctcgtc ttcctcgatc gcgacacgga tcagcccctt gggatagtca
     5161 gcctcggcta ccctgcctac gcggagcaag ttgccgtgac gccgactgtc tcgagtctag
     5221 tcgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5641 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     5701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5821 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtctagt tcagccagct
     5881 cgtcgtgatg tcgaaaccca agccaccctc agaggtgaag gccgcttcga gcacatcggg
     5941 aagcgtgtcg acgacggtgc tcggtgcctc gggtaagagc cagatagatg cggtgacgtt
     6001 caccgtcgcg atcgtggcgg gaaccacggt cactgcgtcc gttacaactc gcacgtcgtc
     6061 ggccagcacg actgactcga cggcttcgat tagcccggga gaggcaaggc catcaggctc
     6121 ggtagacaga atgctgatta acacctcgcc gggagcaggg ctgctcacag ccgcgtcctt
     6181 cacccgtggg tcagccgtca gcgcttgata gcggtaccag gccgcaccgc ctgcggtcga
     6241 gctgcccttg atccgctcga tggtgcgatc gcgaagctcg ccatccggct cattcggctg
     6301 ccgcaccacg ccgtagaacg cggcaaggtt gtcgaggtcg gggccgtttg agta
//
