LOCUS       sequence-102041-        4169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  used to characterize the promoter activities of PBAD for dCas9 and 
            sgRNA response functions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4169)
  AUTHORS   Nielsen AA, Voigt CA
  TITLE     Multi-input CRISPR/Cas genetic circuits that interface host 
            regulatory networks.
  JOURNAL   Mol Syst Biol. 2014 Nov 24;10:763. doi: 10.15252/msb.20145735.
  PUBMED    25422271
REFERENCE   2  (bases 1 to 4169)
  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: "Mol Syst 
            Biol."; date: "2014-11-24"; volume: "10:763. doi"; pages: " 
            10.15252/msb.20145735"
COMMENT     Sequence Label: pAN-PBAD-YFP
FEATURES             Location/Qualifiers
     source          1..4169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    91..112
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(113..991)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1018..1302
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1225..1244
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(1331..1333,1334..1336,1337..2050)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1376..1397)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1637..1656)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1984..2005
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2070..2141
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2157..2184
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2310..2898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2390..2409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3069..3860,3861..3929))
                     /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     3692..3711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3930..4034)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4102..4120
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4124..4167)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 ttcagccaaa aaacttaaga ccgccggtct tgtccactac cttgcagtaa tgcggtggac
       61 aggatcggcg gttttctttt ctcttctcaa gaattcgcgg ccgcttctag agttatgaca
      121 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
      181 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
      241 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
      301 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
      361 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
      421 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
      481 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
      541 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
      601 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
      661 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
      721 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
      781 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
      841 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
      901 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
      961 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
     1021 aaaccaattg tccatattgc atcagacatt gtcgtcactg cgtcttttac tggctcttct
     1081 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     1141 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     1201 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     1261 ctacctgacg ctttttatcg caactctcta ctgtttctcc atatactaga gtcacacagg
     1321 aaagtactag atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1381 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1441 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1501 ctggcccacc ctcgtgacca ccttcggcta cggcctgcaa tgcttcgccc gctaccccga
     1561 ccacatgaag ctgcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1621 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1681 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1741 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1801 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1861 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1921 cgacaaccac tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga
     1981 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2041 gtacaagtaa taatactaga gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2101 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     2161 caccttcggg tgggcctttc tgcgtttata gctcactcaa aggcggtaat acggttatcc
     2221 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     2281 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2341 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2401 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2461 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2521 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2581 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2641 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2701 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2761 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2821 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2881 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2941 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3001 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3061 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3121 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3181 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     3241 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3301 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3361 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3421 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3481 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3541 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3601 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3661 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3721 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3781 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3841 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3901 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4021 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     4081 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
     4141 tccttagctt tcgctaagga tgatttctg
//
