LOCUS       sequence-121218-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Rci gene (driven by the inducible T7 promoter) and 11 
            fragment barcode cassette in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  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 8060)
  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: "Nucleic 
            Acids Res."; date: "2014"; volume: "42(16):e127. doi"; pages: " 
            10.1093/nar/gku604. Epub 2014 Jul 10"
COMMENT     Sequence Label: DIG70 (T7->Rci; 11bc)
FEATURES             Location/Qualifiers
     source          1..8060
                     /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        494..598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(599..667,668..1459)
                     /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(817..836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1630..2218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2119..2138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3901..3918
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3933..4075
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4001..4020)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4161..4183
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4177..4368)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5177..6259)
                     /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     6221..6240
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6260..6337)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6543..6562
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6646..6665
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6646..6664
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6665..6689
                     /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             6704..6726
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7904..7921
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7974..7992)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7988..8035
                     /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 attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 tgcgaacctc atcactcgtt gcgctagcgg caatactttc gtgccaatcc ggtacgtggg
     2341 atataggatt gatttttctt cactgtacgc cggagaacat aagaagcgtc ataaccctat
     2401 cagtcccaag aataagtccc agtattaact gtttcagcac cacgtaccgg attggcacgc
     2461 ttgcagagaa atagggaagg ctcatagcgt tcgaagtagt tcccaaccag aacactcctg
     2521 gtgagtcgct ggaagccatc tatagaaagg acgacgctcg ggccggtgta gccaagcttg
     2581 ttgtgccaat ccggtacgtg gacaacgtca acgttaatct cgtgcatcag atcctagttg
     2641 gttgagaaca gagcgcgacg tggggctggg agccgatacc cgtaagtagg agagggcagc
     2701 cccaggtacc ggattggcac tggcctacgt gcgccatcgc accattaaac atacccggat
     2761 cgatatttag gtgtgcgggg ctcctgcatc aattaataca tcgttcgaac cagacccgtc
     2821 cattcatctc gcctgctggc ctacgtgcca atccggtacg tgggcgatta atcagcatcc
     2881 aaggttgcac accggtttaa tcgtgggaca tcgtcaagcg tcaatatcct aaagacctgc
     2941 gaggtttagg catcagtgcg gggccacgta ccggattggc acgaaagtat tgccacctcg
     3001 aaagtgacga ttgttattga tcggtttgca ccccgaccag ccaggcatct catctccacg
     3061 gggatcgagg gcatcaatca catttgacgg atccggcaat actttcgtgc caatccggta
     3121 cgtggtgtat gcaagggaaa ttaacggacg gcctcaattc ctgcaaggta acgcaccgcg
     3181 gcccaagagc cattaagcga tataatcgca catctggcca acccgccacg taccggattg
     3241 gcacgcttgc agagaatcct gggctcttca accaacagtg acgggggctt attaggagga
     3301 ttttgatacg gacgcgcaac cgtcgtcagg cagctttgaa gcccgtctct ccggatgcca
     3361 agcttgttgt gccaatccgg tacgtggcgg ccgagttcgc tcaccttttt gaatcctcgg
     3421 acgccataac taacagcccg tttatggaag agtatgcact gcctaagggc ggagggccaa
     3481 gagtcctcca ggtaccggat tggcactggc ctacgtgccc caattaccac attaaagata
     3541 tctgcactgg cgtcccctct tctcgaggac gagggtaaaa aagcgccatt gcactaggac
     3601 ttaccgcgga gactgcctct gctggcctac gtgccaatcc ggtacgtggc gttgcatgta
     3661 ttgcagcctc agggacgtca gtggatcatg aaggtagagc atgcgtcctc tgctgttaaa
     3721 atctgagttc tggacaaact accaattggc cacgtaccgg attggcacga aagtattgcc
     3781 ggacagcatc ttccttgcct caacatgttc tttcctgcgt tatcccctga ttctgtggat
     3841 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3901 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3961 ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc tctgatgccg
     4021 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     4081 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     4141 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     4201 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     4261 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     4321 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     4381 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     4441 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     4501 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     4561 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     4621 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     4681 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     4741 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     4801 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     4861 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     4921 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     4981 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     5041 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     5101 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     5161 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5221 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     5281 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     5341 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     5401 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     5461 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     5521 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     5581 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     5641 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     5701 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     5761 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     5821 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     5881 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     5941 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     6001 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     6061 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     6121 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     6181 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     6241 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     6301 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     6361 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     6421 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6481 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6541 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6601 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     6661 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     6721 aggagatata catatgccgt ctccacgcat ccgtaaaatg tccctgtcac gcgcactgga
     6781 taagtacctg aaaacagttt ctgttcacaa gaaagggcat caacaggagt tttaccggag
     6841 caatgttatc aagcgatatc ccattgctct tcggaatatg gacgaaataa caaccgttga
     6901 tattgctaca tacagagacg ttcgtttagc agaaataaac ccccgaacgg gtaaagccat
     6961 tacaggtaat actgtacgtc ttgaactcgc ccttctgtca tctctgttca atattgctcg
     7021 tgttgaatgg ggaacctgtc gtactaaccc ggttgaactg gttcgcaagc cgaaagtatc
     7081 ctccggacga gatcgccggc taacgtcttc agaagaacgt cgcctttctc gctatttccg
     7141 cgaaaaaaat ctgatgttgt atgtcatttt ccatcttgcc cttgaaacag ccatgcggca
     7201 gggcgaaata ctggccttac gttgggagca cattgatttg cgccacggtg tggctcattt
     7261 acctgaaacc aaaaacggtc actcacggga tgttcctctg tccagacgtg cccgtaactt
     7321 tcttcaaatg atgcccgtta atctccacgg caatgttttt gattacaccg catccggctt
     7381 taaaaatgcc tggagaatag ccacacaacg acttcgcatc gaggacctgc attttcacga
     7441 tctacggcat gaagcaataa gccgcttctt cgaactgggt agcctgaatg taatggagat
     7501 tgctgcaata tcaggacatc gttccatgaa tatgctgaaa cggtatactc atcttcgtgc
     7561 atggcaactg gtcagtaagc ttgatgcccg ccggcggcag acacaaaaag tggcagcatg
     7621 gtttgtgccg tatcctgccc atatcacgac tatcgatgaa gaaaatgggc agaaagcgca
     7681 tcgtattgag atcggtgatt ttgataacct tcacgtcact gccacaacaa aagaggaagc
     7741 agttcaccgc gccagtgagg ttttgttgcg tacactggcc attgcagcac agaaaggcga
     7801 acgtgtccca tctcccggag cgttacctgt taacgaccct gactacatta tgatttgccc
     7861 tctgaacccg ggcagcacac cgctgtaagc ggccgcactc gagcaccacc accaccacca
     7921 ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     7981 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     8041 aggaggaact atatccggat
//
