LOCUS       PACE_AP_Neg_pBAD        4503 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  AUTHORS   Nicholas Southern
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4503
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="recombinant plasmid"
     CDS             complement(1006..1197)
                     /label=Rop CDS
                     /note="Rop"
                     /note="color: #993366; direction: LEFT"
     terminator      1308..1803
                     /label=long terminator
                     /note="/created_by=User"
                     /note="color: #ffffff"
     misc_feature    1808..2203
                     /label=Geneious type: Concatenated sequence
                     /label=nonstandard type: Concatenated sequence
                     /note="color: #a6acb3"
     promoter        1881..2165
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli;
                     thearaC regulatory gene is transcribed in the 
                     oppositedirection (Guzman et al., 1995)"
                     /note="color: #ffffff"
     protein_bind    1883..1899
                     /note="araO2"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2020..2036
                     /note="araO1"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2041..2057
                     /note="araO1"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2062..2083
                     /note="CAP"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2094..2110
                     /note="araI1"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2115..2131
                     /note="araI2"
                     /note="color: #31849b; direction: RIGHT"
     RBS             2204..2218
                     /label=SD8 RBS
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     CDS             2219..3313
                     /label=pIII-neg
                     /note="/created_by=User"
                     /note="color: #993366; direction: RIGHT"
     misc_feature    3314..3355
                     /label=tetA/orfL bidirectional terminator
                     /note="termination of bla transcript is slightly weaker
                     than opposite direction
                     "
                     /note="color: #ffffff"
     misc_feature    3356..3368
                     /label=User Junction Modified Base
                     /note="color: #a6acb3"
     terminator      complement(3383..3429)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
                     /note="color: #ffffff; direction: LEFT"
     CDS             complement(3480..4340)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="color: #ccffcc
                     Cleavage site after base 4271"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4341..4445)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      join(4494..4503,1..578)
                     /label=ColE1 origin of replication
                     /note="color: #ffff00"
ORIGIN
        1 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
       61 gtttgccgga tcaagagcta ccacctcttt ttccgaaggt aactggcttc agcagagcgc
      121 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      181 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      241 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
      301 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
      361 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
      421 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      481 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      541 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      601 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
      781 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      841 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
      901 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      961 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     1021 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     1081 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     1141 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
     1201 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
     1261 gatgctcacg atacgggtta ctgatgatga acaagaggac atccggtaag gccaaatagg
     1321 ggttcataag gtacggcccg tactgaaccc gggtaggctg cttgagccag tgagcgattg
     1381 ctggcctaga tgaatgactg tccacgacag aacccggctt atcggtcagt ttcacctgat
     1441 ttacgtaaaa acccgcttcg gcgggttttt gcttttggag gggcagaaag atgaatgact
     1501 gtccacgacg ctatacccaa aagaaagcgg cttatcggtc agtttcacct ggtttacgta
     1561 aaaacccgct tcggcgggtt tttgcttttg gaggggcaga aagatgaatg actgtccacg
     1621 acactatacc caaaagaaag cggcttatcg gtcagtttca cctgttttac gtaaaaaccc
     1681 gcttcggcgg gtttttactt ttggaggggc agaaagatga atgactgtcc acgacactat
     1741 acccaaaaga aagcggctta tcggtcagtt ttacctgatg tacgtaataa accgttccgg
     1801 cgggtttatg cctttaatta aggagttttg caggtgcctt ggaacacctg cttttcgctg
     1861 aattcgcggc cgcttctaga aagaaaccaa ttgtccatat tgcatcagac attgccgtca
     1921 ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat
     1981 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
     2041 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
     2101 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
     2161 tccatgaccg aattcaaaag atcttttaag aaggagatat acaaaggagg aaaaaaaaat
     2221 gaaaaaatta ttattcgcaa ttcctttagt tgttcctttc tattctcact ccgctgaaac
     2281 tgttcatcac catcaccatc acgctgaaac tgttgaaagt tgtttagcaa aaccccatac
     2341 agaaaattca tttactaacg tctggaaaga cgacaaaact ttagatcgtt acgctaacta
     2401 tgagggctgt ctgtggaatg ctacaggcgt tgtagtttgt actggtgacg aaactcagtg
     2461 ttacggtaca tgggttccta ttgggcttgc tatccctgaa aatgagggtg gtggctctga
     2521 gggtggcggt tctgagggtg gcggttctga gggtggcggt actaaacctc ctgagtacgg
     2581 tgatacacct attccgggct atacttatat caaccctctc gacggcactt atccgcctgg
     2641 tactgagcaa aaccccgcta atcctaatcc ttctcttgag gagtctcagc ctcttaatac
     2701 tttcatgttt cagaataata ggttccgaaa taggcagggg gcattaactg tttatacggg
     2761 cactgttact caaggcactg accccgttaa aacttattac cagtacactc ctgtatcatc
     2821 aaaagccatg tatgacgctt actggaacgg taaattcaga gactgcgctt tccattctgg
     2881 ctttaatgag gatccattcg tttgtgaata tcaaggccaa tcgtctgacc tgcctcaacc
     2941 tcctgtcaat gctggcggcg gctctggtgg tggttctggt ggcggctctg agggtggtgg
     3001 ctctgagggt ggcggttctg agggtggcgg ctctgaggga ggcggttccg gtggtggctc
     3061 ttcccaaatg gctcaagtcg gtgacggtga taattcacct ttaatgaata atttccgtca
     3121 atatttacct tccctccctc aatcggttga atgtcgccct tttgtctttg gcgctggtaa
     3181 accttacgag ttcagtatcg actgcgataa gatcaacctg ttccgcggtg tctttgcgtt
     3241 tcttttatat gttgccacct ttatgtatgt attttctacg tttgctaaca tactgcgtaa
     3301 taaggagtct taaaaatata atgaccctct tgataaccca agagggcatt ttttaatgcc
     3361 catggcgtag ctgtgctcta gaataaaacg aaaggcccag tctttcgact gagcctttcg
     3421 ttttatttga ccggaaatca gaattggtta attggttgta acactggcag agcattacgt
     3481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4441 ccgcgattaa ttgcgttgcg ctcactgccc gctttccagg ccaaataggc cgttgagatc
     4501 ctt
//
