LOCUS       sequence-200259-        7264 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses Rci gene (driven by the inducible T7 promoter) and 5 
            fragment barcode cassette in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7264)
  AUTHORS   Peikon ID, Gizatullina DI, Zador AM
  TITLE     In vivo generation of DNA sequence diversity for cellular barcoding.
  JOURNAL   Nucleic Acids Res. 2014;42(16):e127. doi: 10.1093/nar/gku604. Epub 
            2014 Jul 10.
  PUBMED    25013177
REFERENCE   2  (bases 1 to 7264)
  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: "Nucleic 
            Acids Res."; date: "2014"; volume: "42(16):e127. doi"; pages: " 
            10.1093/nar/gku604. Epub 2014 Jul 10"
COMMENT     Sequence Label: IDP205 (T7->Rci; 5bc)
FEATURES             Location/Qualifiers
     source          1..7264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3108..3125
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3140..3279
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3205..3224)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3365..3387
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3381..3572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4381..5463)
                     /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     5425..5444
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5464..5541)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5747..5766
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5850..5869
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5850..5868
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5869..5893
                     /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             5908..5930
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7108..7125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7178..7196)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7192..7239
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttt
     2281 gcgaacctca tcactcgttg cggcaatact ttcgtgccaa tccggtacgt ggtgtatgca
     2341 agggaaatta acggacggcc tcaattcctg caaggtaacg caccgcggcc caagagccat
     2401 taagcgatat aatcgcacat ctggccaacc cgccacgtac cggattggca cgcttgcaga
     2461 gaatcctggg ctcttcaacc aacagtgacg ggggcttatt aggaggattt tgatacggac
     2521 gcgcaaccgt cgtcaggcag ctttgaagcc cgtctctccg gatgccaagc ttgttgtgcc
     2581 aatccggtac gtggcggccg agttcgctca cctttttgaa tcctcggacg ccataactaa
     2641 cagcccgttt atggaagagt atgcactgcc taagggcgga gggccaagag tcctccaggt
     2701 accggattgg cactggccta cgtgccccaa ttaccacatt aaagatatct gcactggcgt
     2761 cccctcttct cgaggacgag ggtaaaaaag cgccattgca ctaggactta ccgcggagac
     2821 tgcctctgct ggcctacgtg ccaatccggt acgtggcgtt gcatgtattg cagcctcagg
     2881 gacgtcagtg gatcatgaag gtagagcatg cgtcctctgc tgttaaaatc tgagttctgg
     2941 acaaactacc aattggccac gtaccggatt ggcacgaaag tattgccgga cagcatcttc
     3001 cttgcctcaa catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3061 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3121 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3181 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3241 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3301 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3361 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3421 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     3481 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     3541 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     3601 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     3661 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     3721 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     3781 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     3841 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     3901 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     3961 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     4021 acgatcatgc gcacccgtgg ggccgccatg ccggcgataa tggcctgctt ctcgccgaaa
     4081 cgtttggtgg cgggaccagt gacgaaggct tgagcgaggg cgtgcaagat tccgaatacc
     4141 gcaagcgaca ggccgatcat cgtcgcgctc cagcgaaagc ggtcctcgcc gaaaatgacc
     4201 cagagcgctg ccggcacctg tcctacgagt tgcatgataa agaagacagt cataagtgcg
     4261 gcgacgatag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     4321 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     4381 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4441 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4501 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4561 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4621 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atatccgcac caacgcgcag
     4681 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4741 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4801 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg
     4861 ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat
     4921 ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga
     4981 gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt
     5041 agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag
     5101 cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct
     5161 tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc
     5221 cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa
     5281 cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat
     5341 cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg
     5401 ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt
     5461 cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5521 tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     5581 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     5641 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     5701 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     5761 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     5821 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     5881 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
     5941 ccgtctccac gcatccgtaa aatgtccctg tcacgcgcac tggataagta cctgaaaaca
     6001 gtttctgttc acaagaaagg gcatcaacag gagttttacc ggagcaatgt tatcaagcga
     6061 tatcccattg ctcttcggaa tatggacgaa ataacaaccg ttgatattgc tacatacaga
     6121 gacgttcgtt tagcagaaat aaacccccga acgggtaaag ccattacagg taatactgta
     6181 cgtcttgaac tcgcccttct gtcatctctg ttcaatattg ctcgtgttga atggggaacc
     6241 tgtcgtacta acccggttga actggttcgc aagccgaaag tatcctccgg acgagatcgc
     6301 cggctaacgt cttcagaaga acgtcgcctt tctcgctatt tccgcgaaaa aaatctgatg
     6361 ttgtatgtca ttttccatct tgcccttgaa acagccatgc ggcagggcga aatactggcc
     6421 ttacgttggg agcacattga tttgcgccac ggtgtggctc atttacctga aaccaaaaac
     6481 ggtcactcac gggatgttcc tctgtccaga cgtgcccgta actttcttca aatgatgccc
     6541 gttaatctcc acggcaatgt ttttgattac accgcatccg gctttaaaaa tgcctggaga
     6601 atagccacac aacgacttcg catcgaggac ctgcattttc acgatctacg gcatgaagca
     6661 ataagccgct tcttcgaact gggtagcctg aatgtaatgg agattgctgc aatatcagga
     6721 catcgttcca tgaatatgct gaaacggtat actcatcttc gtgcatggca actggtcagt
     6781 aagcttgatg cccgccggcg gcagacacaa aaagtggcag catggtttgt gccgtatcct
     6841 gcccatatca cgactatcga tgaagaaaat gggcagaaag cgcatcgtat tgagatcggt
     6901 gattttgata accttcacgt cactgccaca acaaaagagg aagcagttca ccgcgccagt
     6961 gaggttttgt tgcgtacact ggccattgca gcacagaaag gcgaacgtgt cccatctccc
     7021 ggagcgttac ctgttaacga ccctgactac attatgattt gccctctgaa cccgggcagc
     7081 acaccgctgt aagcggccgc actcgagcac caccaccacc accactgaga tccggctgct
     7141 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     7201 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     7261 ggat
//
