LOCUS       sequence-417388-        5734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even ORI-marker screen destination vector; contains sfYFP dropout 
            for barcoded-ORI insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 4254 to 4288)
  AUTHORS   Gilbert C, Lee H
  TITLE     POSSUM Expansion Pack
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5734)
  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: 
            "unpublished"
COMMENT     Sequence Label: pGL2_211
FEATURES             Location/Qualifiers
     source          1..5734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(485..579)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(600..1259)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1174..1193
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1260..1362)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1449..1471
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1463..1480
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1464..1480
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1780..1791
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1798..2511
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             2809..4185
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     promoter        4254..4288
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     oriT            5214..5322
                     /note="incP origin of transfer"
     primer_bind     complement(5476..5492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5476..5492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5489..5511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      join(5682..5734,1..336)
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
ORIGIN
        1 gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac cttaaaggct
       61 ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga ggctatttaa
      121 gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt gaaacatgag
      181 agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta gttcgttaaa
      241 catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag agcttagtac
      301 gtactatcaa caggttgaac tgctgatctt cagatcggcg cgccggccgg ccgaagatcc
      361 tttgatcttt tctacggggt ctgacgctct attcaacaaa gccgccgtcc cgtcaagtca
      421 gcgtaaatgg gtagggggct tcaaatcgtc cccccatacg atataagttg ttactagtgc
      481 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
      541 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc gatatcaaat
      601 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
      661 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
      721 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
      781 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgagacgaa aaacatattc
      841 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
      901 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
      961 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     1021 ccgtctttca ttgccatacg aaattccgga tgagcattca tcaggcgggc aagaatgtga
     1081 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     1141 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     1201 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     1261 ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg tagtgatctt
     1321 atttcattat ggtgaaagtt ggaacctctt acgtgccgat caacgtctca ttgatacctg
     1381 aaacaaaacc catcgtacgg ccaaggaagt ctccaataac tgtgatccac cacaagcgcc
     1441 agggttttcc cagtcacgac gttgtaaaac gacggccagt catgcataat ccgcacgcat
     1501 ctggaataag gaagtgccat tccgcctgac cttagcgtgc gaacagcacg ctattgttgt
     1561 atcattactc gcatccattc tcaggctgtc tcgtctcgtc tccgaggtct caggagaatg
     1621 tgaagagctt cgaaagtgaa acgtgatttc atgcgtcatt ttgaacattt tgtaaatctt
     1681 atttaataat gtgtgcggca attcacattt aatttatgaa tgttttctta acatcgcggc
     1741 aactcaagaa acggcaggtt cggatcttag ctactagaga aagaggagaa atactagatg
     1801 cgtaaaggcg aagaactgtt cactggtgtc gtccctattc tggtggaact ggatggtgat
     1861 gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac taatggtaaa
     1921 ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc gactctggta
     1981 acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat gaagcagcat
     2041 gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat ttcctttaag
     2101 gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac cctggtaaac
     2161 cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg ccataagctg
     2221 gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa aaatggcatt
     2281 aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct ggctgatcac
     2341 taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa tcactatctg
     2401 agctaccaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat ggttctgctg
     2461 gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa ataattcagc
     2521 caaaaaactt aagaccgccg gtcttgtcca ctaccttgca gtaatgcggt ggacaggatc
     2581 ggcggttttc ttttctcttc tcaagaagct cttcagcaag gagggatttt atgaccgatg
     2641 atgaagaaaa gaatctgaaa cttagtttat atgtggtaaa atgttttaat caagtttagg
     2701 aggaattaat tatgaagtgt aattaatgta acagggttca attaaaagag ggaagcgtat
     2761 cattaaccct ataaactacg tctgccctca ttattggagg gtgaaataat gaatacatcc
     2821 tattcacaat cgaatttacg acacaaccaa attttaattt ggctttgcat tttatctttt
     2881 tttagcgtat taaatgaaat ggttttgaac gtcagcttac ctgatattgc aaatgatttt
     2941 aataaacctc ctgcgagtac aaactgggtg aacacagcct ttatgttaac cttttccatt
     3001 ggaacagctg tatatggaaa gctatctgat caattaggca tcaaaaggtt actcctattt
     3061 ggaattataa taaattgttt cgggtcggta attgggtttg ttggccattc tttcttttcc
     3121 ttacttatta tggctcgttt tattcaaggg gctggtgcag ctgcatttcc agcactcgta
     3181 atggttgtag ttgcgcgcta tattccaaag gaaaataggg gtaaagcatt tggtcttatt
     3241 ggatcgatag tagccatggg agaaggagtc ggtccagcga ttggtggaat gatagcccat
     3301 tatattcatt ggtcctatct tctactcatt cctatgataa caattatcac tgttccgttt
     3361 cttatgaaat tattaaagaa agaagtaagg ataaaaggtc attttgatat caaaggaatt
     3421 atactaatgt ctgtaggcat tgtatttttt atgttgttta caacatcata tagcatttct
     3481 tttcttatcg ttagcgtgct gtcattcctg atatttgtaa aacatatcag gaaagtaaca
     3541 gatccttttg ttgatcccgg attagggaaa aatatacctt ttatgattgg agttctttgt
     3601 gggggaatta tatttggaac agtagcaggg tttgtctcta tggttcctta tatgatgaaa
     3661 gatgttcacc agctaagtac tgccgaaatc ggaagtgtaa ttattttccc tggaacaatg
     3721 agtgtcatta ttttcggcta cattggtggg atacttgttg atagaagagg tcctttatac
     3781 gtgttaaaca tcggagttac atttctttct gttagctttt taactgcttc ctttctttta
     3841 gaaacaacat catggttcat gacaattata atcgtatttg ttttaggtgg gctttcgttc
     3901 accaaaacag ttatatcaac aattgtttca agtagcttga aacagcagga agctggtgct
     3961 ggaatgagtt tgcttaactt taccagcttt ttatcagagg gaacaggtat tgcaattgta
     4021 ggtggtttat tatccatacc cttacttgat caaaggttgt tacctatgga agttgatcag
     4081 tcaacttatc tgtatagtaa tttgttatta cttttttcag gaatcattgt cattagttgg
     4141 ctggttacct tgaatgtata taaacattct caaagggatt tctaagctta tcgttaaggg
     4201 atcaactttg ggagagagtt caaaattgat ccttttttta cgctatacag gagttgacgg
     4261 ctagctcagt cctaggtaca gtgctagcta ctaggaggaa aaaaatggtg agtaagggtg
     4321 aagccgtgat taaagagttc atgcgcttca aagtgcatat ggagggctca atgaacggcc
     4381 atgaatttga aattgaaggt gaaggagaag gccgtccgta tgagggcacc cagaccgcca
     4441 aactgaaagt cacaaagggt ggcccgctgc cgttttcgtg ggatatttta tcccctcaat
     4501 tcatgtacgg cagccgggcg tttattaaac acccggcgga tatcccggat tactataagc
     4561 aatcttttcc agaaggcttt aaatgggagc gcgtgatgaa ttttgaagac ggaggtgctg
     4621 ttaccgtgac ccaggacacg agcttagaag acggtacctt gatctataag gtaaagctgc
     4681 gtggtaccaa tttcccgccg gatgggccag ttatgcagaa aaagactatg ggctgggaag
     4741 catcgacgga gcgtctttac cccgaagacg gcgtcctgaa aggcgatatt aaaatggccc
     4801 tgcgactgaa agacggcggt cgctaccttg cagatttcaa aacaacgtat aaagcgaaaa
     4861 aacctgtgca gatgcccggg gcgtacaacg ttgatcgcaa actagatatc acttcacaca
     4921 acgaagatta cacggtagtc gaacagtatg aacgtagcga ggggcgccat agcaccggag
     4981 gcatggatga actctataaa taagcttcgc tcgtgctcgc cgcgaccttc ctcttcatgc
     5041 acctcttcgg catcgggctg cacaacgcta cagaggagct tttccgctgc ataaccctgc
     5101 ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg atatacagga
     5161 ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca gccgggcagg
     5221 ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc ttattcgcac
     5281 ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtacgctag gtatacttga
     5341 gaccacccca ggatacatag attaccacaa ctccgagccc ttccacctag cgtgctaacc
     5401 acgcacgcta ttgttgtata ggctaggtgg aggctcagtg atgataagtc tgcgatggtg
     5461 gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcagagggca
     5521 caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc agttcggcgt
     5581 atggcatatg tcgctggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5641 aaaaaggccg cgttgctggc gttacagctg ggcgcgcccc atgtcagccg ttaagtgttc
     5701 ctgtgtcact caaaattgct ttgagaggct ctaa
//
