LOCUS       sequence-298656-        4214 bp    DNA     circular SYN 15-DEC-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 Dec 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"
     protein_bind    5..21
                     /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             59..70
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    797..817
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     promoter        864..941
                     /gene="lacI"
                     /label=lacI promoter
     CDS             942..2024
                     /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(961..980)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(2222..2239)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2393..2981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2473..2492)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3305..3964)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3879..3898
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     4126..4144
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4148..4191)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4194..4214,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggaattgtga gcggataaca attttgacgg tgtgagcgga taacagtaca gtggatccaa
       61 agaggagaaa aagcttatgc gtaaaggcga agaactgttc acgggcgtag ttccgattct
      121 ggtcgagctg gacggcgatg tgaacggtca taagtttagc gttcgcggtg aaggtgaggg
      181 cgacgcgacc aacggcaaac tgaccctgaa gttcatctgc accaccggta aactgccggt
      241 gccttggccg accttggtga cgacgttgac gtatggcgtg cagtgttttg cgcgttatcc
      301 ggaccacatg aaacaacacg atttcttcaa atctgcgatg ccggagggtt acgtccagga
      361 gcgtaccatt tccttcaagg atgatggcta ctacaaaact cgcgcagagg ttaagtttga
      421 aggtgacacg ctggtcaatc gtatcgaatt gaagggtatc gactttaaag aggatggtaa
      481 cattctgggc cataaactgg agtataactt caacagccat aatgtttaca ttacggcaga
      541 caagcaaaag aacggcatca aggccaattt caagattcgc cacaatgttg aggacggtag
      601 cgtccaactg gccgaccatt accagcagaa caccccaatt ggtgacggtc cggttttgct
      661 gccggataat cactatctga gcacccaaag cgtgctgagc aaagatccga acgaaaaacg
      721 tgatcacatg gtcctgctgg aatttgtgac cgctgcgggc atcacccacg gtatggacga
      781 gctgtataag taatgatact agtagcggcc gctgcagtgc ttcctaatgc aggagtcgca
      841 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
      901 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
      961 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     1021 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     1081 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     1141 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     1201 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     1261 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     1321 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     1381 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     1441 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     1501 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     1561 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     1621 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     1681 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     1741 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     1801 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     1861 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     1921 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     1981 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     2041 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     2101 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgagatct
     2161 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2221 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2281 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2341 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2401 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2461 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2521 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2581 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2641 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2701 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2761 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2821 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2881 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2941 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3001 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3061 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3121 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3181 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3241 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagtcga
     3301 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     3361 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     3421 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     3481 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca
     3541 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     3601 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     3661 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     3721 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     3781 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     3841 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     3901 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     3961 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     4021 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaac tcgagtgcca
     4081 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4141 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     4201 cggccgcttc taga
//
