LOCUS       sequence-306377-        4214 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of sfGFP under Synthetic promoter (SynP) and cloning of 
            the sgRNA with simple screening of positive clone by fluorescence .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4214)
  AUTHORS   Tan SI, Yu PJ, Ng IS
  TITLE     CRISPRi-mediated Programming Essential Gene can as a Direct 
  JOURNAL   Biotechnol Bioeng. 2020 May 27. doi: 10.1002/bit.27443.
  PUBMED    32458463
REFERENCE   2  (bases 1 to 4214)
  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: "Biotechnol 
            Bioeng. 2020 May 27. doi: 10.1002/bit.27443."
COMMENT     Sequence Label: pSUI-SynP-sfGFP
FEATURES             Location/Qualifiers
     source          1..4214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    721..741
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        788..865
                     /gene="lacI"
                     /label=lacI promoter
     CDS             866..1948
                     /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(885..904)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(2146..2163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2317..2905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2397..2416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3229..3888)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3803..3822
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4050..4068
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4072..4115)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4118..4139
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    4143..4159
                     /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)."
     RBS             4197..4208
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 atgcgtaaag gcgaagaact gttcacgggc gtagttccga ttctggtcga gctggacggc
       61 gatgtgaacg gtcataagtt tagcgttcgc ggtgaaggtg agggcgacgc gaccaacggc
      121 aaactgaccc tgaagttcat ctgcaccacc ggtaaactgc cggtgccttg gccgaccttg
      181 gtgacgacgt tgacgtatgg cgtgcagtgt tttgcgcgtt atccggacca catgaaacaa
      241 cacgatttct tcaaatctgc gatgccggag ggttacgtcc aggagcgtac catttccttc
      301 aaggatgatg gctactacaa aactcgcgca gaggttaagt ttgaaggtga cacgctggtc
      361 aatcgtatcg aattgaaggg tatcgacttt aaagaggatg gtaacattct gggccataaa
      421 ctggagtata acttcaacag ccataatgtt tacattacgg cagacaagca aaagaacggc
      481 atcaaggcca atttcaagat tcgccacaat gttgaggacg gtagcgtcca actggccgac
      541 cattaccagc agaacacccc aattggtgac ggtccggttt tgctgccgga taatcactat
      601 ctgagcaccc aaagcgtgct gagcaaagat ccgaacgaaa aacgtgatca catggtcctg
      661 ctggaatttg tgaccgctgc gggcatcacc cacggtatgg acgagctgta taagtaatga
      721 tactagtagc ggccgctgca gtgcttccta atgcaggagt cgcataaggg agagcgtcga
      781 gatcccggac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
      841 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
      901 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
      961 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     1021 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     1081 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     1141 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     1201 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     1261 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     1321 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     1381 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     1441 ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc
     1501 gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat
     1561 gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg
     1621 cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg
     1681 atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga
     1741 ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc
     1801 ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc
     1861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     1921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc
     1981 attaggcacc gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     2041 ggtgggcgcg gggcatgact atcgtcgccg cacttatgag atctattaat gaatcggcca
     2101 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2161 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2221 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2281 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2341 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2401 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2461 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2521 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2581 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2641 aagacacgac ttatcgccac cggcagcagc cactggtaac aggattagca gagcgaggta
     2701 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2761 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2821 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2881 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2941 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3001 cacctagatc ctcttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3061 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3121 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3181 cttaccatct ggctccagtg ctgcaatgat accgcgagag tcgacaaatt acgccccgcc
     3241 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     3301 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     3361 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     3421 aaaactggtg aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc
     3481 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     3541 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     3601 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     3661 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     3721 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     3781 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     3841 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     3901 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     3961 gtgaaagttg gaacctctta cgtgcccgat caactcgagt gccacctgac gtctaagaaa
     4021 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat
     4081 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagagg
     4141 aattgtgagc ggataacaat tttgacggtg tgagcggata acagtacagt ggatccaaag
     4201 aggagaaaaa gctt
//
