LOCUS       sequence-183388-        9023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ORF part assembly entry vector derived from pHT01. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9023)
  AUTHORS   Gilbert C, Howarth M, Harwood CR, Ellis T
  TITLE     Extracellular Self-Assembly of Functional and Tunable Protein 
            Conjugates from Bacillus subtilis.
  JOURNAL   ACS Synth Biol. 2017 Mar 7. doi: 10.1021/acssynbio.6b00292.
  PUBMED    28230977
REFERENCE   2  (bases 1 to 9023)
  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: "ACS Synth 
            Biol. 2017 Mar 7. doi: 10.1021/acssynbio.6b00292."
COMMENT     Sequence Label: pCG004
FEATURES             Location/Qualifiers
     source          1..9023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        293..397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(398..466,467..1258)
                     /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     complement(616..635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1429..2017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1918..1937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2171..2188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3437..3459
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3451..3468
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3452..3468
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3511..4593)
                     /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
                     EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4555..4574
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    4977..5001
                     /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)."
     CDS             5180..5893
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5210..5238)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5833..5853
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      5916..5987
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6003..6030
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 tttttatgta gcaatatatc cgatacagca ccttctaaaa gcgttggtga atagggcatt
       61 ttacctatct cctctcattt tgtggaataa aaatagtcat attcgtccat ctacctatcc
      121 tattatcgaa cagttgaact ttttaatcaa ggatcagtcc tttttttcat tattcttaaa
      181 ctgtgctctt aactttaaca actcgatttg tttttccaga tctcgagggt aactagcctc
      241 gccgatcccg caagaggccc ggcagtcagg tggcactttt cggggaaatg tgcgcggaac
      301 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
      361 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      421 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      481 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      541 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      601 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      661 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      721 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      781 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
      841 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
      901 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
      961 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1021 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1081 tattgctgat aaatctggag ccggtgagcg tggttctcgc ggtatcattg cagcactggg
     1141 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     1201 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1261 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1321 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     1381 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1441 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1501 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1561 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1621 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1681 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1741 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1801 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1861 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1921 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1981 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     2041 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     2101 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     2161 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcatgcttaa
     2221 gttattggta tgactggttt taagcgcaaa aaaagttgct ttttcgtacc tattaatgta
     2281 tcgttagaaa accgactgta aaaagtacag tcggcattat ctcatattat aaaagccagt
     2341 cattaggcct atctgacaat tcctgaatag agttcataaa caatcctgca tgataaccat
     2401 cacaaacaga atgatgtacc tgtaaagata gcggtaaata tattgaatta cctttattaa
     2461 tgaattttcc tgctgtaata atgggtagaa ggtaattact attattattg atatttaagt
     2521 taaacccagt aaatgaagtc catggaataa tagaaagaga aaaagcattt tcaggtatag
     2581 gtgttttggg aaacaatttc cccgaaccat tatatttctc tacatcagaa aggtataaat
     2641 cataaaactc tttgaagtca ttctttacag gagtccaaat accagagaat gttttagata
     2701 caccatcaaa aattgtataa agtggctcta acttatccca ataacctaac tctccgtcgc
     2761 tattgtaacc agttctaaaa gctgtatttg agtttatcac ccttgtcact aagaaaataa
     2821 atgcagggta aaatttatat ccttcttgtt ttatgtttcg gtataaaaca ctaatatcaa
     2881 tttctgtggt tatactaaaa gtcgtttgtt ggttcaaata atgattaaat atctcttttc
     2941 tcttccaatt gtctaaatca attttattaa agttcatttg atatgcctcc taaattttta
     3001 tctaaagtga atttaggagg cttacttgtc tgctttcttc attagaatca atcctttttt
     3061 aaaagtcaat attactgtaa cataaatata tattttaaaa atatcccact ttatccaatt
     3121 ttcgtttgtt gaactaatgg gtgctttagt tgaagaataa aagaccacat taaaaaatgt
     3181 ggtcttttgt gtttttttaa aggatttgag cgtagcgaaa aatccttttc tttcttatct
     3241 tgataataag ggtaactatt gccgtcgtcc attccgacag catcgccagt cactatggcg
     3301 tgctgctagc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     3361 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     3421 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attcgagctc
     3481 aggccttaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3541 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3601 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3661 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3721 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     3781 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     3841 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     3901 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     3961 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4021 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4081 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4141 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4201 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4261 caccgccgtt ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4321 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4381 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4441 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4501 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4561 ctctgcgaca tcgtataacg ttactggttt catcaaaatc gtctccctcc gtttgaatat
     4621 ttgattgatc gtaaccagat gaagcactct ttccactatc cctacagtgt tatggcttga
     4681 acaatcacga aacaataatt ggtacgtacg atctttcagc cgactcaaac atcaaatctt
     4741 acaaatgtag tctttgaaag tattacatat gtaagattta aatgcaaccg ttttttcgga
     4801 aggaaatgat gacctcgttt ccaccggaat tagcttggta ccagctattg taacataatc
     4861 ggtacggggg tgaaaaagct aacggaaaag ggagcggaaa agaatgatgt aagcgtgaaa
     4921 aattttttat cttatcactt gaaattggaa gggagattct ttattataag aattgtggaa
     4981 ttgtgagcgg ataacaattc ccaattaaag gaggaagggg atagagaccg ggcctttctg
     5041 cgtttatata ctagagaatt ttgtcaaaat aattttattg acaacgtctt attaacgttg
     5101 atataattta aattttattt gacaaaaatg ggctcgtgtt gtacaataaa tgttactaga
     5161 gaaaggtggt gaatactaga tgcgtaaagg agaagaactt ttcactggag ttgtcccaat
     5221 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     5281 aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg gaaaactacc
     5341 tgttccatgg ccaacacttg tcactacttt cggttatggt gttcaatgct ttgcgagata
     5401 cccagatcat atgaaacagc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     5461 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtgctg aagtcaagtt
     5521 tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta aagaagatgg
     5581 aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat acatcatggc
     5641 agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
     5701 aagcgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     5761 tttaccagac aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa
     5821 gagagatcac atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga
     5881 tgaactatac aaataataat actagagcca ggcatcaaat aaaacgaaag gctcagtcga
     5941 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     6001 ctggctcacc ttcgggtggg cctttctgcg tttatatact agagatccgt ttaggctggg
     6061 cggtgatagc ttctcgttca ggcagtaggt ctcagttcta agacgtcccc ggggcagccc
     6121 gcctaatgag cgggcttttt tcacgtcacg cgtccatgga gatctttgtc tgcaactgaa
     6181 aagtttatac cttacctgga acaaatggtt gaaacatacg aggctaatat cggcttatta
     6241 ggaatagtcc ctgtactaat aaaatcaggt ggatcagttg atcagtatat tttggacgaa
     6301 gctcggaaag aatttggaga tgacttgctt aattccacaa ttaaattaag ggaaagaata
     6361 aagcgatttg atgttcaagg aatcacggaa gaagatactc atgataaaga agctctaaaa
     6421 ctattcaata accttacaat ggaattgatc gaaagggtgg aaggttaatg gtacgaaaat
     6481 taggggatct acctagaaag ccacaaggcg ataggtcaag cttaaagaac ccttacatgg
     6541 atcttacaga ttctgaaagt aaagaaacaa cagaggttaa acaaacagaa ccaaaaagaa
     6601 aaaaagcatt gttgaaaaca atgaaagttg atgtttcaat ccataataag attaaatcgc
     6661 tgcacgaaat tctggcagca tccgaaggga attcatatta cttagaggat actattgaga
     6721 gagctattga taagatggtt gagacattac ctgagagcca aaaaactttt tatgaatatg
     6781 aattaaaaaa aagaaccaac aaaggctgag acagactcca aacgagtctg tttttttaaa
     6841 aaaaatatta ggagcattga atatatatta gagaattaag aaagacatgg gaataaaaat
     6901 attttaaatc cagtaaaaat atgataagat tatttcagaa tatgaagaac tctgtttgtt
     6961 tttgatgaaa aaacaaacaa aaaaaatcca cctaacggaa tctcaattta actaacagcg
     7021 gccaaactga gaagttaaat ttgagaaggg gaaaaggcgg atttatactt gtatttaact
     7081 atctccattt taacatttta ttaaacccca tacaagtgaa aatcctcttt tacactgttc
     7141 ctttaggtga tcgcggaggg acattatgag tgaagtaaac ctaaaaggaa atacagatga
     7201 attagtgtat tatcgacagc aaaccactgg aaataaaatc gccaggaaga gaatcaaaaa
     7261 agggaaagaa gaagtttatt atgttgctga aacggaagag aagatatgga cagaagagca
     7321 aataaaaaac ttttctttag acaaatttgg tacgcatata ccttacatag aaggtcatta
     7381 tacaatctta aataattact tctttgattt ttggggctat tttttaggtg ctgaaggaat
     7441 tgcgctctat gctcacctaa ctcgttatgc atacggcagc aaagactttt gctttcctag
     7501 tctacaaaca atcgctaaaa aaatggacaa gactcctgtt acagttagag gctacttgaa
     7561 actgcttgaa aggtacggtt ttatttggaa ggtaaacgtc cgtaataaaa ccaaggataa
     7621 cacagaggaa tccccgattt ttaagattag acgtaaggtt cctttgcttt cagaagaact
     7681 tttaaatgga aaccctaata ttgaaattcc agatgacgag gaagcacatg taaagaaggc
     7741 tttaaaaaag gaaaaagagg gtcttccaaa ggttttgaaa aaagagcacg atgaatttgt
     7801 taaaaaaatg atggatgagt cagaaacaat taatattcca gaggccttac aatatgacac
     7861 aatgtatgaa gatatactca gtaaaggaga aattcgaaaa gaaatcaaaa aacaaatacc
     7921 taatcctaca acatcttttg agagtatatc aatgacaact gaagaggaaa aagtcgacag
     7981 tactttaaaa agcgaaatgc aaaatcgtgt ctctaagcct tcttttgata cctggtttaa
     8041 aaacactaag atcaaaattg aaaataaaaa ttgtttatta cttgtaccga gtgaatttgc
     8101 atttgaatgg attaagaaaa gatatttaga aacaattaaa acagtccttg aagaagctgg
     8161 atatgttttc gaaaaaatcg aactaagaaa agtgcaataa actgctgaag tatttcagca
     8221 gtttttttta tttagaaata gtgaaaaaaa tataatcagg gaggtatcaa tatttaatga
     8281 gtactgattt aaatttattt agactggaat taataattaa cacgtagact aattaaaatt
     8341 taatgaggga taaagaggat acaaaaatat taatttcaat ccctattaaa ttttaacaag
     8401 ggggggatta aaatttaatt agaggtttat ccacaagaaa agaccctaat aaaattttta
     8461 ctagggttat aacactgatt aatttcttaa tgggggaggg attaaaattt aatgacaaag
     8521 aaaacaatct tttaagaaaa gcttttaaaa gataataata aaaagagctt tgcgattaag
     8581 caaaactctt tactttttca ttgacattat caaattcatc gatttcaaat tgttgttgta
     8641 tcataaagtt aattctgttt tgcacaacct tttcaggaat ataaaacaca tctgaggctt
     8701 gttttataaa ctcagggtcg ctaaagtcaa tgtaacgtag catatgatat ggtatagctt
     8761 ccacccaagt tagcctttct gcttcttctg aatgtttttc atatacttcc atgggtatct
     8821 ctaaatgatt ttcctcatgt agcaaggtat gagcaaaaag tttatggaat tgatagttcc
     8881 tctctttttc ttcaactttt ttatctaaaa caaacacttt aacatctgag tcaatgtaag
     8941 cataagatgt ttttccagtc ataatttcaa tcccaaatct tttagacaga aattctggac
     9001 gtaaatcttt tggtgaaaga att
//
