LOCUS       sequence-396453-        6182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose induced expression of repL(AT2) gene, which triggers DNA 
            replication in trans, weak level of constitutive GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6182)
  AUTHORS   Huan YW, Brown R, Wang B
  TITLE     An adenine/thymidine-rich region is integral to RepL-mediated DNA 
            replication.
  JOURNAL   Front Microbiol. 2023 Feb 9;14:1095671. doi: 
            10.3389/fmicb.2023.1095671. eCollection 2023.
  PUBMED    36846746
REFERENCE   2  (bases 1 to 6182)
  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: "Front 
            Microbiol."; date: "2023-02-9"; volume: "14:1095671. doi"; pages: " 
            10.3389/fmicb.2023.1095671. eCollection 2023"
COMMENT     Sequence Label: 4A3.PBAD-repL(AT2).J23115-gfp
FEATURES             Location/Qualifiers
     source          1..6182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..42)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(58..129)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(152..865)
                     /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(192..212)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     807..835
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     RBS             872..883
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(895..929)
                     /label=BBa_J23115
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.15"
     CDS             complement(981..1859)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1886..2170
                     /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     2093..2112
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(3361..4311)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4359..4581
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(5073..5167)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(5197..5988,5989..6057))
                     /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     5820..5839
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6058..6162)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgtcta agaacgcaga aaggcccacc cgaaggtgag ccagtgtgac tctagtagag
       61 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
      121 ttttatttga tgcctggctc tagtattatt atttgtatag ttcatccatg ccatgtgtaa
      181 tcccagcagc tgttacaaac tcaagaagga ccatgtggtc tctcttttcg ttgggatctt
      241 tcgaaagggc agattgtgtg gacaggtaat ggttgtctgg taaaaggaca gggccatcgc
      301 caattggagt attttgttga taatggtctg ctagttgaac gcttccatct tcaatgttgt
      361 gtctaatttt gaagttaact ttgattccat tcttttgttt gtctgccatg atgtatacat
      421 tgtgtgagtt atagttgtat tccaatttgt gtccaagaat gtttccatct tctttaaaat
      481 caataccttt taactcgatt ctattaacaa gggtatcacc ttcaaacttg acttcagcac
      541 gtgtcttgta gttcccgtca tctttgaaaa atatagttct ttcctgtaca taaccttcgg
      601 gcatggcact cttgaaaaag tcatgctgtt tcatatgatc tgggtatctc gcaaagcatt
      661 gaacaccata accgaaagta gtgacaagtg ttggccatgg aacaggtagt tttccagtag
      721 tgcaaataaa tttaagggta agttttccgt atgttgcatc accttcaccc tctccactga
      781 cagaaaattt gtgcccatta acatcaccat ctaattcaac aagaattggg acaactccag
      841 tgaaaagttc ttctccttta cgcatctagt atttctcctc tttaatctct agtagctagc
      901 attgtaccaa gggctgagct agctataaat tttgttatca ataaaaaagg ccgcgatttg
      961 cggccttatt gttcgtctta ttatgacaac ttgacggcta catcattcac tttttcttca
     1021 caaccggcac gaaactcgct cgggctggcc ccggtgcatt ttttaaatac tcgcgagaaa
     1081 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtagtg
     1141 ctcaaaagca gcttcgcctg actaatgcgt tggtcctcgc gccagcttaa gacgctaatc
     1201 cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     1261 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     1321 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt
     1381 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     1441 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     1501 aagaaacccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     1561 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     1621 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcagacaaa ttctcgtccc
     1681 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     1741 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     1801 accagatggg cgttaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata
     1861 cttttcatac tcccaccatt cagagaagaa accaattgtc catattgcat cagacattgc
     1921 cgtcactgcg tcttttactg gctcttctcg ctaacccaac cggtaacccc gcttattaaa
     1981 agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat
     2041 aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata
     2101 gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca actctctact
     2161 gtttctccat gtacctgtcg cggcaacgcg ctaacagacg tagtaagaac caccagcatt
     2221 gtaatgctgg ctaaagtcac tttcctgagc tgtataacga tgagcgattt tactttttct
     2281 ggctatgaat tggcctgctt tgtaacacac tccggtctat cccgtagcgc cgggcatatc
     2341 ctgtcgcaat gtgcaaatct cgcggcaaca accagtgaat acttcattca caagcctcac
     2401 cgcctgatcg cggcagaaac tggttatagc caatcaaccg tcgttcgtgc attccgtgaa
     2461 gctgtaaaca aaggaattct gtctgtagag attgttatcg gcgatcaccg tgaacgtcgc
     2521 gctaacctgt accggtttac accatccttt ttggccttcg cacaacaagc caaaaatgcg
     2581 ctgatagaaa gcaaattaaa gatctcttca gcggcaacca aggttaaagc tgttctcgct
     2641 aagacattgg ctttatttaa ttttttatcc acacccccat gtcaaaatga taccccctcc
     2701 ccctgtcagg atgacgtggc gatcaagaac aagaagagcc aggtgaagaa gacgaagcgt
     2761 agcgtttccg gcggtgccgg aacaaccagc ctcaaaaaat tgacttcatg gatcgctaag
     2821 gcaaaagcaa aggctgacaa tctgcggtta tccaaaaaac gcactcaaaa acatgagttc
     2881 aagcagaaag tagaggcggc tgcgcggaaa tatgcttacc tgaagaacaa gcgttcgcct
     2941 gatattggcg ggatatcaaa cttcgataac ctaccgcatt gcatgacggt aaacgaagct
     3001 cttaatgcgg ttttagccaa aaataaagat aacgaacaat ggggtatacc ggcaggattc
     3061 agagggtaat aactcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt
     3121 ttcgttttgg tccgctcact caaaggcggt aatctcgagg ttacattgtc gatctgttca
     3181 tggtgaacag ctttgaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt
     3241 acaccgtttt catctgtgca tatggacagt tttccctttg atatgtaacg gtgaacagtt
     3301 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
     3361 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
     3421 ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc
     3481 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
     3541 atgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
     3601 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
     3661 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
     3721 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
     3781 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
     3841 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
     3901 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
     3961 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
     4021 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
     4081 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
     4141 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
     4201 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
     4261 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
     4321 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattaca
     4381 tgtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
     4441 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
     4501 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
     4561 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
     4621 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
     4681 cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga
     4741 gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg
     4801 gtaaatggca ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag
     4861 aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct
     4921 gactttttgc tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta
     4981 tcccgtgaca ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca
     5041 ggcttacccg tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg
     5101 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac
     5161 aggagtccaa gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5221 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5281 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5341 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5401 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5461 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5521 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5581 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5641 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5701 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5761 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5821 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5881 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5941 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6001 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6061 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6121 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc ccatggtgcc
     6181 ac
//
