LOCUS       sequence-201976-        7355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PBAD-TP901 and Ptet-Bxb1 as well as constitutive TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7355)
  AUTHORS   Hsiao V, Hori Y, Rothemund PW, Murray RM
  TITLE     A population-based temporal logic gate for timing and recording 
            chemical events.
  JOURNAL   Mol Syst Biol. 2016 May 17;12(5):869. doi: 10.15252/msb.20156663.
  PUBMED    27193783
REFERENCE   2  (bases 1 to 7355)
  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: "2016-05-17"; volume: "12(5):869. doi"; pages: " 
            10.15252/msb.20156663"
COMMENT     Sequence Label: pVHed05
FEATURES             Location/Qualifiers
     source          1..7355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..22
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(23..649)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGPELIICGLEKQLKCESGS"
     primer_bind     554..573
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        665..720
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    701..719
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    761..779
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    786..804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             849..2348
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="RALVVIRLSRVTDATTSPERQLESCQQLCAQRGWDVVGVAEDLDV
                     SGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLVV
                     SATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGYL
                     PTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQGR
                     EPQGREWSATALKRSMISGAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAEL
                     VKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTVA
                     MAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLIGSPAYRAGS
                     PQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSMN
                     VRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      2377..2404
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2487..3365)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWAYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3392..3676
                     /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     3599..3618
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3730..3747
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5215..5247
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag
                     /note="wild-type LAA variant"
                     /translation="AAKDENYALAA"
     misc_feature    5251..5271
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5272..5329
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5353..5370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5580..6167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5660..5679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6255..6349)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6370..7029)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTIQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6944..6963
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(7030..7132)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7309..7353)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 gaattcgcgg ccgcttctag agttaagacc cactttcaca tttaagttgt ttttctaatc
       61 cgcatatgat caattcaggg ccgaataaga aggctggctc tgcaccttgg tgatcaaata
      121 attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt tccctttctt
      181 ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa tgccccacag
      241 cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa aaggctaatt
      301 gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt acttttgctc
      361 catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt aaaaaatctt
      421 gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac atctcaatgg
      481 ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta ggctgctcta
      541 cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc tcattaagca
      601 gctctaatgc gctgttaatc actttacttt tatctaatct tgacatcatt aattcctaat
      661 ttttgttgac actctatcgt tgatagagtt attttaccac tccctatcag tgatagagaa
      721 aagtgaaaat ccatatgact agcagatcca ctagagtcga tccctatcag tgatagagat
      781 tgacatccct atcagtgata gagatactga gcacaagctt gcggctttca cacataggag
      841 ctagcgtgag agccctggta gtcatccgcc tgtcccgcgt caccgatgct acgacttcac
      901 cggagcgtca gctggagtct tgccagcagc tctgcgccca gcgcggctgg gacgtcgtcg
      961 gggtagcgga ggatctggac gtctccgggg cggtcgatcc gttcgaccgg aagcgcagac
     1021 cgaacctggc ccggtggcta gcgttcgagg agcaaccgtt tgacgtgatc gtggcgtacc
     1081 gggtagatcg gttgacccga tcgatccggc atcttcagca gctggtccac tgggccgagg
     1141 accacaagaa gctggtcgtc tccgcgaccg aagcgcactt cgatacgacg acgccgtttg
     1201 cggcggtcgt catcgcgctt atgggaacgg tggcgcagat ggaattagaa gcgatcaaag
     1261 agcggaaccg ttcggctgcg catttcaata tccgcgccgg gaaataccga ggatccctgc
     1321 cgccgtgggg atacctgcct acgcgcgtgg acggggagtg gcggctggtg ccggaccctg
     1381 tgcagcgaga gcgcatcctc gaggtgtatc accgcgtcgt cgacaaccac gagccgctgc
     1441 atctggtggc ccacgacctg aaccggcgtg gtgtcctgtc gccgaaggac tacttcgcgc
     1501 agctgcaagg ccgcgagccg cagggccggg agtggtcggc taccgcgctg aagcgatcga
     1561 tgatctccgg ggcgatgctc gggtacgcga ctctgaacgg taagaccgtc cgagacgacg
     1621 acggagcccc gctggtgcgg gctgagccga tcctgacccg tgagcagctg gaggcgctgc
     1681 gcgccgagct cgtgaagacc tcccgggcga agcccgcggt gtctaccccg tcgctgctgc
     1741 tgcgggtgtt gttctgcgcg gtgtgcgggg agcccgcgta caagttcgcc gggggaggac
     1801 gtaagcaccc gcgctaccgc tgccgctcga tggggttccc gaagcactgc gggaacggca
     1861 cggtggcgat ggccgagtgg gacgcgttct gcgaggagca ggtactggat ctgctcgggg
     1921 acgcggagcg tctggagaaa gtctgggtag cgggctcgga ctccgcggtc gaactcgcgg
     1981 aggtgaacgc ggagctggtg gacctgacgt cgctgatcgg ctccccggcc taccgggcgg
     2041 gctctccgca gcgagaagca ctggatgccc gtattgcggc gctggccgcg cggcaagagg
     2101 agctggaggg cctggaggct cgcccgtctg gctgggagtg gcgcgagacc gggcagcggt
     2161 tcggggactg gtggcgggag caggacaccg cggcaaagaa cacctggctt cggtcgatga
     2221 acgttcggct gacgttcgac gtccgcggcg ggctgactcg cacgatcgac ttcggggatc
     2281 ttcaggagta cgagcagcat ctcaggctcg gcagcgtggt cgaacggcta cacaccggga
     2341 tgtcgtaata atactagatg cattaataat cacactggct caccttcggg tgggcctttc
     2401 tgcgtttata tactagagag agaatataaa aagccagatt attaatccgg cttttttatt
     2461 atttcctgca gggcggccgc catatgttat gacaacttga cggctacatc attcactttt
     2521 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     2581 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     2641 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     2701 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     2761 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     2821 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     2881 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     2941 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     3001 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     3061 gcgcgcggac gaaagtaagc ccactggtga taccattcgc gagcctccgg atgacgaccg
     3121 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     3181 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     3241 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     3301 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     3361 gccatacttt tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga
     3421 cattgccgtc actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt
     3481 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     3541 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac tctttgctat
     3601 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     3661 tctactgttt ctccataccg tttttttggg ctagctacta gagtcacaca ggaaacctac
     3721 tagatgaaac atcatcacca tcaccaccag gccggcacta agaaagtagc aatctataca
     3781 cgagtatcca ctactaacca agcagaggaa ggcttctcaa ttgatgagca aattgaccgt
     3841 ttaacaaaat atgctgaagc aatggggtgg caagtatctg atacttatac tgatgctggt
     3901 ttttcagggg ccaaacttga acgcccagca atgcaaagat taatcaacga tatcgagaat
     3961 aaagcttttg atacagttct tgtatataag ctagaccgcc tttcacgtag tgtaagagat
     4021 actctttatc ttgttaagga tgtgttcaca aaaaataaaa tagactttat ctcgcttaat
     4081 gaaagtattg atacttcttc tgctatgggt agcttgtttc tcactattct ttctgcaatt
     4141 aatgagtttg aaagagagaa tataaaagaa cgcatgacta tgggtaaact agggcgagcg
     4201 aaatctggta agtctatgat gtggactaag acagcttttg ggtattacca caacagaaag
     4261 acaggtatat tagaaattgt tcctttacaa gctacaatag ttgaacaaat attcactgat
     4321 tatttatcag gaatatcact tacaaaatta agagataaac tcaatgaatc tggacacatc
     4381 ggtaaagata taccgtggtc ttatcgtacc ctaagacaaa cacttgataa tccagtttac
     4441 tgtggttata tcaaatttaa ggacagccta tttgaaggta tgcacaaacc aattatccct
     4501 tatgagactt atttaaaagt tcaaaaagag ctagaagaaa gacaacagca gacttatgaa
     4561 agaaataaca accctagacc tttccaagct aaatatatgc tgtcagggat ggcaaggtgc
     4621 ggttactgtg gagcaccttt aaaaattgtt cttggccaca aaagaaaaga tggaagccgc
     4681 actatgaaat atcactgtgc aaatagattt cctcgaaaaa caaaaggaat tacagtatat
     4741 aatgacaata aaaagtgtga ttcaggaact tatgatttaa gtaatttaga aaatactgtt
     4801 attgacaacc tgattggatt tcaagaaaat aatgactcct tattgaaaat tatcaatggc
     4861 aacaaccaac ctattcttga tacttcgtca tttaaaaagc aaatttcaca gatcgataaa
     4921 aaaatacaaa agaactctga tttgtaccta aatgatttta tcactatgga tgagttgaaa
     4981 gatcgtactg attcccttca ggctgagaaa aagctgctta aagctaagat tagcgaaaat
     5041 aaatttaatg actctactga tgtttttgag ttagttaaaa ctcagttggg ctcaattccg
     5101 attaatgaac tatcatatga taataaaaag aaaatcgtca acaaccttgt atcaaaggtt
     5161 gatgttactg ctgataatgt agatatcata tttaaattcc aactcgctac cggtgctgct
     5221 aaggacgaaa actacgctct ggctgcttaa tactagtagc ggccgctgca gtccggcaaa
     5281 aaagggcaag gtgtcaccac cctgcccttt ttctttaaaa ccgaaaagat tacttcgcgt
     5341 tatgcaggct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5401 atcagctcac tcaaaggcgg taatcctagc tagggcgttc ggctgcggcg agcggtatca
     5461 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5521 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5581 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5641 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5701 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5761 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5821 aagctgggct gtatgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5881 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5941 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6001 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6061 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6241 tgacttgtgc ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac
     6301 aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc
     6361 gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat
     6421 tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag
     6481 caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc
     6541 catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa
     6601 aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac
     6661 atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga
     6721 tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat
     6781 caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc
     6841 aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa
     6901 ggccgtaata tccagctgaa tggtctggtt ataggtacat tgagcaactg actgaaatgc
     6961 ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt
     7021 tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     7081 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca
     7141 ttttcgccag atatcgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7201 aggcgtatca cgaggccctt tcgtcttcac ctcgagtgcc acctgacgtc taagaaacca
     7261 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa
     7321 aaaaaatcct tagctttcgc taaggatgat ttctg
//
