LOCUS       sequence-372118-        5808 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mCherry under T7 promoter control.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5808)
  AUTHORS   Baumschlager A, Aoki SK, Khammash M
  TITLE     Dynamic Blue Light-Inducible T7 RNA Polymerases (Opto-T7RNAPs) for 
            Precise Spatiotemporal Gene Expression Control.
  JOURNAL   ACS Synth Biol. 2017 Nov 17;6(11):2157-2167. doi: 
            10.1021/acssynbio.7b00169. Epub 2017 Oct 18.
  PUBMED    29045151
REFERENCE   2  (bases 1 to 5808)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.7b00169"; journalName: "ACS Synth Biol"; date: 
            "2017-11-17- 17"; volume: "6"; issue: "11"; pages: "2157-2167"
COMMENT     Sequence Label: pAB50
FEATURES             Location/Qualifiers
     source          1..5808
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(23..100)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     306..325
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1178..1222
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1261..1279)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1275..1322
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1366..1821
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     1663..1684
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1936..2727,2728..2796))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2559..2578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2797..2889)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2970..3558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3459..3478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3712..3729
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3744..3883
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3809..3828)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3969..3991
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3985..4176)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4748..5808,1..22))
                     /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     join(5792..5808,1..3)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
       61 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
      121 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
      181 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
      241 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
      301 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
      361 gccacgatgc gtccggcgta gcctaggtaa tacgactcac tatagggaga ggatcctgtc
      421 aatttccgcg atagaggagg taaagcatat ggtttcaaaa ggcgaagaag acaacatggc
      481 gattatcaag gaatttatgc gtttcaaggt ccacatggaa ggcagcgtca atggtcacga
      541 atttgaaatt gaaggcgaag gtgaaggccg tccgtatgaa ggcacccaga cggcaaaact
      601 gaaggtcacc aaaggcggtc cgctgccgtt tgcttgggat attctgtcac cgcaattcat
      661 gtatggttcg aaagcgtacg ttaagcatcc ggccgatatc ccggactatc tgaaactgtc
      721 ctttccggaa ggcttcaaat gggaacgtgt tatgaacttc gaagatggcg gtgtggttac
      781 cgtcacgcag gatagctctc tgcaagacgg tgaatttatt tataaagtga agctgcgcgg
      841 caccaatttc ccgagcgatg gtccggttat gcagaaaaag acgatgggct gggaagcgag
      901 ttccgaacgt atgtacccgg aagacggtgc cctgaaaggc gaaatcaagc agcgcctgaa
      961 actgaaggat ggcggtcact atgacgcaga agtgaaaacc acgtacaagg ctaaaaagcc
     1021 ggtccaactg ccgggtgcat acaacgtgaa catcaagctg gatatcacca gccataacga
     1081 agactatacg atcgttgaac agtacgaacg tgcagaaggc cgccactcta ccggcggtat
     1141 ggatgaactg tacaaataag gtaccctcga gtctggtaaa gaaaccgctg ctgcgaaatt
     1201 tgaacgccag cacatggact cgtctactag tcgcagctta attaacctaa actgctgcca
     1261 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1321 tgctagcgaa aggaggagtc gactatatcc ggattggcga atgggacgcg ccctgtagcg
     1381 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1441 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1501 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1561 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1621 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1681 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1741 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1801 aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt
     1861 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1921 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1981 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2041 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2101 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2161 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2221 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2281 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2341 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2401 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2461 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2521 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2581 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2641 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2701 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2761 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt
     2821 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     2881 ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3601 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3661 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3721 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacaca tctgtgcggt
     3781 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3841 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3901 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3961 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4021 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     4081 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     4141 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     4201 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     4261 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     4321 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     4381 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     4441 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatat
     4501 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     4561 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     4621 gagcacgatc atgctagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     4681 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     4741 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4801 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
     4861 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
     4921 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
     4981 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg tccgcaccaa
     5041 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
     5101 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
     5161 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
     5221 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
     5281 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
     5341 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
     5401 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
     5461 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
     5521 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
     5581 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
     5641 tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
     5701 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca
     5761 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacat
//
